3,4-dihydro-2,7-naphthyridine-1,6(2h,7h)-diones as mek inhibitors

ABSTRACT

The invention relates to a method of treating a MEK-associated tumor by administering to a subject in need thereof a therapeutically effective amount of a solid form 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.

FIELD OF THE INVENTION

The present invention relates to novel3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione compounds, orpharmaceutically acceptable salts thereof, which act as MEK inhibitorsand are useful for the treatment of abnormal cell growth, such ascancer, in patients. The present invention also relates topharmaceutical compositions containing the compounds and to methods ofusing the compounds and compositions in the treatment of abnormal cellgrowth, such as cancer, in subject in need thereof. The presentinvention also relates to solid forms of8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,to pharmaceutical compositions containing the solid forms, and tomethods of using the solid forms and compositions thereof in thetreatment of abnormal cell growth, such as cancer, in subject in needthereof.

BACKGROUND OF THE INVENTION

MEK kinase (Mitogen Activated Protein Kinase Kinase (MAPKK)) is animportant component of the Ras-RAF-MEK-ERK cell survival pathway. TheRas pathway is activated by binding of growth factors, cytokines, andhormones to their cognate receptors. In cancer cells, this pathway is,however, constitutively activated and leads to increased cancer cellsurvival, cell proliferation, angiogenesis, and metastasis. The tumorsthat show constitutive activation of the pathway include, but are notlimited to, those of the colon, pancreas, breast, brain, ovary, lungs,and skin. Activation of Ras (due to upstream signaling or as a result ofactivating point mutations in the Ras oncogene) lead to thephosphorylation and activation of Raf kinase that in turn phosphorylateand activate MEK1 and MEK2 (also known as MAPKK1 and MAPKK2). MEK1 andMEK2 are dual-specificity kinases that activate ERK1 and ERK2 byphosphorylating and activating the ERK1/2 kinase (also referred to asMAP Kinase) that further phosphorylates and regulates the function ofproteins such as Mcl-1, Bim and Bad that are involved in cell survivaland apoptosis. Thus, activation of this phosphorylation mediated cascadeleads to enhanced cell proliferation, cell survival, and decreased celldeath that are necessary for initiation and maintenance of thetumorigenic phenotype. Inhibition of this pathway, particularlyinhibiting MEK activity, is known to be beneficial in treatinghyperproliferative diseases. MEK inhibitors have shown variable degreesof activity in several settings, including BRAF V600-mutant melanoma,NRAS-mutant melanoma, low-grade serous ovarian cancer, plexiformneurofibromas, thyroid cancer, and low-grade gliomas, with more limitedresponses in KRAS-mutant pancreatic cancer or lung cancer.

Cancers that frequently metastasize to the brain, e.g., melanoma andnon-small cell lung cancer, are known to carry MAPK pathway activatingalterations such as the BRAF V600E and KRAS G12 mutation (Cancer GenomeAtlas N., Cell 2015; 161:1681-96). Although activating mutations canoccur at different levels in the canonical pathway, they all requiresignaling via mitogen/extracellular signal-regulated kinase (MEK) inorder to increase proliferation and survival (Schubbert S, Shannon K,Bollag G., Nat Rev Cancer. 2007; 7:295-308). The common activation ofthe MAPK pathway in malignancies and the central and downstream locationof MEK also render MEK inhibitors of potential interest for thetreatment of intracranial tumors.

Blood-brain interfaces comprise the cerebral microvessel endotheliumforming the blood-brain barrier (BBB) and the epithelium of the choroidplexuses forming the blood-CSF barrier (BCSFB). The blood brain barrier(BBB) is a highly selective physical, transport and metabolic barrierthat divides the CNS from the blood. The BBB may prevent certain drugsfrom entering brain tissue and is therefore a limiting factor in thedelivery of many peripherally-administered agents to the CNS. Theefficacy of many molecularly targeted agents in central nervous systemtumors is limited by penetration across the blood-brain barrier (BBB),which is composed of a monolayer of endothelial cells connected by tightjunctions that serve as a physical barrier protecting the brain. Inaddition, these endothelial cells express multiple efflux transporters,including P-glycoprotein (P-gp) and breast cancer resistance protein(BCRP), which are known to exclude many anticancer agents from the brain(Ohtsuki and Terasaki, 2007, Pharm Res 24:1745-1758; Agarwal et al.,2011, Pharm Res 24:1745-1758). Similar to the blood-brain barrier, theblood-CSF barrier functions to prevent the passage of most blood-bornesubstances into the brain, while selectively permitting the passagespecific substances into the brain and facilitating the removal of brainmetabolites and metabolic products into the blood.

Thus, there remains a need for therapies for the treatment of tumorsmediated by MEK, including therapies that can penetrate the BBB and/orBCSFB and target tumors in the CNS.

SUMMARY OF THE INVENTION

Provided herein, in part, are compounds of Formula I and Formula II andpharmaceutically acceptable salts thereof. Such compounds can inhibitthe activity of MEK, thereby effecting biological functions, and may beuseful for treating a subject having a MEK-associated tumor. Alsoprovided herein are solid forms of8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Also provided are pharmaceutical compositions and medicaments comprisingthe compounds according to any of the formulae described herein, andpharmaceutically acceptable salts thereof, which may be useful fortreating a subject having a MEK-associated tumor alone or in combinationwith additional anticancer therapies. Also provided herein are methodsfor preparing the compounds, pharmaceutically acceptable salts, andpharmaceutical compositions according to any of the formulae describedherein and pharmaceutically acceptable salts thereof, and methods ofusing the foregoing. This summary is provided to introduce a selectionof concepts in a simplified form that are further described below in thedetailed description. This summary is not intended to identify keyfeatures or essential features of the claimed subject matter, nor is itintended to be used in isolation as an aid in determining the scope ofthe claimed subject matter.

According to an embodiment of the invention, provided herein is acompound of Formula I

or a pharmaceutically acceptable salt thereof, wherein:

-   -   R¹ is H, Br, C1-C6 alkyl or phenyl;    -   R² is H, halogen or CH₃—;    -   R³ is H, hydroxyC1-C6 alkyl-, hydroxyC1-C6 alkoxy-, C1-C6        alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, or (C3-C6        cycloalkyl)C1-C6 alkoxy-; and    -   R⁴ is phenyl substituted with 1, 2 or 3 substituents        independently selected from halogen, C1-C6 alkyl, C1-C6        alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6        alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, and C1-C6        alkyl-C(═O)—.

Also provided herein is a compound of Formula II

or a pharmaceutically acceptable salt thereof, wherein:

-   -   R¹ is H, Br, C1-C6 alkyl or phenyl;    -   R² is H, halogen or CH₃—;    -   R³ is H, hydroxyC1-C6 alkyl-, hydroxyC1-C6 alkoxy-, C1-C6        alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, or (C3-C6        cycloalkyl)C1-C6 alkoxy-; and    -   R^(a) and R^(b) are independently selected from halogen, C1-C6        alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6        alkyl, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, and        C1-C6 alkyl-C(═O)—.

In one embodiment, provided herein are solid forms of8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.

In one embodiment, provided herein is a pharmaceutical compositioncomprising a compound according to any of the formulae described herein,or a pharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier or excipient. In some embodiments, the pharmaceuticalcomposition comprises two or more pharmaceutically acceptable carriersand/or excipients.

In one embodiment, provided herein are therapeutic methods and usescomprising administering a compound according to any of the formulaedescribed herein, or a pharmaceutically acceptable salt thereof, to asubject.

In one embodiment, provided herein is a method for the treatment ofabnormal cell growth, for example a tumor, for example a MEK-associatedtumor, in a subject in need thereof, comprising administering to thesubject a therapeutically effective amount of a compound according toany of the formulae described herein, or a pharmaceutically acceptablesalt thereof. Compounds according to any of the formulae describedherein may be administered as single agents or may be administered incombination with one or more anticancer therapies.

In one embodiment, provided herein is a method for the treatment ofabnormal cell growth, for example a tumor, for example a MEK-associatedtumor, in a subject in need thereof, comprising administering to thesubject an amount of a compound according to any of the formulaedescribed herein, or a pharmaceutically acceptable salt thereof, incombination with an amount of an additional anticancer agent, whichamounts are together effective in treating said abnormal cell growth.

In one embodiment, provided herein is a compound according to any of theformulae described herein, or a pharmaceutically acceptable saltthereof, for use as a medicament.

In one embodiment, provided herein is a compound according to any of theformulae described herein, or a pharmaceutically acceptable saltthereof, for use in the treatment of abnormal cell growth, for example atumor, for example a MEK-associated tumor.

In one embodiment, provided herein is the use of a compound according toany of the formulae described herein, or a pharmaceutically acceptablesalt thereof, for the manufacture of a medicament for the treatment ofabnormal cell growth, for example a tumor, for example a MEK-associatedtumor, in a subject.

In one embodiment, provided herein is a pharmaceutical compositioncomprising a compound according to any of the formulae described herein,or a pharmaceutically acceptable salt thereof, and at least onepharmaceutically acceptable carrier or excipient.

Each of the embodiments of the compounds according to any of theformulae described herein can be combined with one or more otherembodiments of the compounds according to any of the formulae describedherein not inconsistent with the embodiment(s) with which it iscombined.

It is to be understood that both the foregoing general description andthe following detailed description are exemplary and explanatory onlyand are not restrictive of the invention, as claimed.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts a powder X-ray diffraction pattern of crystallineanhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 1.

FIG. 2 . depicts a powder X-ray diffraction pattern of crystallineanhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2.

FIG. 3 . depicts a powder X-ray diffraction pattern of crystallinemonohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3.

FIG. 4 . depicts a powder X-ray diffraction pattern of amorphous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 4.

FIG. 5 depicts a sorption isotherm of crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3.

DETAILED DESCRIPTION OF THE INVENTION

In one aspect, the invention provides a compound of the Formula I

or a pharmaceutically acceptable salt thereof, wherein:

-   -   R¹ is H, Br, C1-C6 alkyl or phenyl;    -   R² is H, halogen or CH₃—;    -   R³ is H, hydroxyC1-C6 alkyl-, hydroxyC1-C6 alkoxy-, C1-C6        alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, or (C3-C6        cycloalkyl)C1-C6 alkoxy-; and    -   R⁴ is phenyl substituted with 1, 2 or 3 substituents        independently selected from halogen, C1-C6 alkyl, C1-C6        alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6        alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, and C1-C6        alkyl-C(═O)—.

As used herein, the singular forms “a”, “an”, and “the” when referringto substituents include plural references unless indicated otherwise.For example, “a” substituent includes one or more substituents.

For complex chemical names employed herein, a substituent group istypically named before the group to which it attaches. For example,methoxyethyl comprises an ethyl backbone with a methoxy substituent.

The term “halogen” means —F (sometimes referred to herein as “fluoro” or“fluoros”), —Cl, —Br and —I.

The term “C1-C6 alkyl” as used herein refers to a saturated linear orbranched-chain monovalent hydrocarbon radical of one to six carbonatoms. Examples of alkyl groups include, but are not limited to, methyl,ethyl, 1-propyl, isopropyl, 1-butyl, isobutyl, sec-butyl, tert-butyl,2-methyl-2-propyl, pentyl, neopentyl, and hexyl.

The term “hydroxyC1-C6 alkyl-” as used herein refers to refers to aC1-C6 alkyl radical as defined herein, wherein one of the hydrogen atomsis replaced with a hydroxy group.

The term “hydroxy” refers to an —OH group.

The term “C3-C6 cycloalkyl” means a fully saturated carbocyclic ringhaving from 3-6 ring carbon atoms. Examples include cyclopropyl,cyclobutyl, cyclopentyl and cyclohexyl.

The term “fluoroC1-C6 alkyl” as used herein refers to a C1-C6 alkylradical as defined herein, wherein one, two or three hydrogen atoms isreplaced with one, two or three fluoro atoms, respectively. Examplesinclude, but are not limited to, fluoromethyl, difluoromethyl,trifluoromethyl, 2-fluoroethyl, 2,2-difluoroethyl, and2,2,2-trifluoroethyl.

The term “C1-C6 alkoxy” as used herein refers to a C1-C6 alkyl radicalas defined herein that is single bonded to an oxygen atom, wherein theradical is on the oxygen atom (i.e., C1-C6-O—). Examples of alkoxygroups include, but are not limited to, methoxy, ethoxy, propoxy, andisopropoxy.

The term “fluoroC1-C6 alkoxy” as used herein refers to a C1-C6 alkoxy asdefined herein, wherein one, two or three hydrogen atoms is replacedwith one, two or three fluoro atoms, respectively. An example includes,but is not limited to, trifluoromethoxy.

The term “(C3-C6 cycloalkyl)C1-C6 alkoxy-” refers to a C1-C6 alkoxy- asdefined herein, wherein one of the hydrogen atoms is replaced with aC3-C6 cycloalkyl group as defined herein.

The term “C1-C6 alkylthio” as used herein refers to a (C1-C6 alkyl)S—radical wherein the C1-C6 alkyl portion is as defined herein.

The term “fluoroC1-C6 alkylthio” as used herein refers to a C1-C6alkylthio group as defined herein, wherein one, two or three hydrogenatoms is replaced with one, two or three fluoro atoms, respectively.

In one embodiment of Formula I, R¹ is H.

In one embodiment of Formula I, R¹ is Br.

In one embodiment of Formula I, R¹ is C1-C6 alkyl. In one embodiment ofFormula I, R¹ is methyl.

In one embodiment of Formula I, R¹ is phenyl.

In one embodiment of Formula I, R² is H.

In one embodiment, R² is halogen.

In one embodiment of Formula I, R² is F.

In one embodiment of Formula I, R² is Cl.

In one embodiment of Formula I, R² is Br.

In one embodiment of Formula I, R² is I.

In one embodiment of Formula I, R² is CH₃—.

In one embodiment of Formula I, R¹ is H and R² is H.

In one embodiment of Formula I, R³ is H.

In one embodiment of Formula I, R³ is hydroxyC1-C6 alkyl-. Nonlimitingexamples include 2-hydroxyethyl.

In one embodiment of Formula I, R³ is hydroxyC1-C6 alkoxy-. Nonlimitingexamples include 2-hydroxyethoxy and 2-hydroxypropoxy having thestructures:

-   -   respectively.

In one embodiment of Formula I, R³ is C1-C6 alkoxy. Nonlimiting examplesinclude methoxy, ethoxy, 1-methylethoxy, and 2,2-dimethylethoxy.

In one embodiment of Formula I, R³ is fluoroC1-C6 alkoxy. A nonlimitingexample includes 2,2-difluoroethoxy.

In one embodiment of Formula I, R³ is C3-C6 cycloalkyl. A nonlimitingexample is cyclopropyl.

In one embodiment of Formula I, R³ is (C3-C6 cycloalkyl)C1-C6 alkoxy-. Anonlimiting example is cyclopropylmethoxy.

In one embodiment of Formula I, R⁴ is phenyl substituted with 1, 2 or 3substituents independently selected from fluoro, chloro, bromo, iodo,ethyl, propyl, isopropyl, methylthio, difluoromethylthio,trifluoromethyl, methoxy, difluoromethoxy, cyclopropyl, and C1-C6alkyl-C(═O)—.

In one embodiment of Formula I, R⁴ is phenyl substituted with 1 or 2substituents independently selected from halogen, C1-C6 alkyl, C1-C6alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6 alkoxy,fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, and C1-C6 alkyl-C(═O)—.

In one embodiment of Formula I, R⁴ is phenyl substituted with 1 or 2substituents independently selected from fluoro, chloro, bromo, iodo,ethyl, propyl, isopropyl, methylthio, difluoromethylthio,trifluoromethyl, methoxy, difluoromethoxy, cyclopropyl, and C1-C6alkyl-C(═O)—.

In one embodiment of Formula I, R⁴ is phenyl substituted with 1substituent selected from halogen, C1-C6 alkyl, C1-C6 alkylthio,fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6 alkoxy, fluoroC1-C6alkoxy, C3-C6 cycloalkyl, and C1-C6 alkyl-C(═O)—.

In one embodiment of Formula I, R⁴ is phenyl substituted with 1substituent selected from fluoro, chloro, bromo, iodo, ethyl, propyl,isopropyl, methylthio, difluoromethylthio, trifluoromethyl, methoxy,difluoromethoxy, cyclopropyl, and C1-C6 alkyl-C(═O)—.

In one embodiment of Formula I, R⁴ is selected from the structures:

In one embodiment of Formula I, R⁴ is

In one embodiment of Formula I, R⁴ is

In one embodiment of Formula I, R⁴ is selected from the structures:

In one embodiment of Formula I, R⁴ is selected from the structures:

In one embodiment, R⁴ has the structure:

wherein R^(a) and R^(b) are independently selected from halogen, C1-C6alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, and C1-C6 alkyl-C(═O)—. Inone embodiment, wherein R^(a) is halogen. In one embodiment, R^(b) ishalogen, C1-C6 alkyl, C1-C6 alkylthio, or fluoroC1-C6 alkoxy. In oneembodiment, wherein R^(a) is halogen and R^(b) is halogen, C1-C6 alkyl,C1-C6 alkylthio, or fluoroC1-C6 alkoxy.

In one embodiment, provided herein is a compound of Formula II:

-   -   or a pharmaceutically acceptable salt thereof, wherein:    -   R¹ is H, Br, C1-C6 alkyl or phenyl;    -   R² is H, halogen or CH₃—;    -   R³ is H, hydroxyC1-C6 alkyl-, hydroxyC1-C6 alkoxy-, C1-C6        alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, or (C3-C6        cycloalkyl)C1-C6 alkoxy-; and    -   R^(a) and R^(b) are independently selected from halogen, C1-C6        alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6        alkyl, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, and        C1-C6 alkyl-C(═O)—.

In one embodiment of Formula II, R¹ is H.

In one embodiment of Formula II, R¹ is Br.

In one embodiment of Formula II, R¹ is C1-C6 alkyl. In one embodiment ofFormula II, R¹ is methyl.

In one embodiment of Formula II, R¹ is phenyl.

In one embodiment of Formula II, R² is H.

In one embodiment of Formula II, R² is halogen.

In one embodiment of Formula II, R² is F.

In one embodiment of Formula II, R² is Cl.

In one embodiment of Formula II, R² is Br.

In one embodiment of Formula II, R² is I.

In one embodiment of Formula II, R² is CH₃—.

In one embodiment of Formula II, R² is H or CH₃—.

In one embodiment of Formula II, R¹ is H and R² is H or CH₃—.

In one embodiment of Formula II, R¹ is H and R² is H.

In one embodiment of Formula II, R³ is H.

In one embodiment of Formula II, R³ is hydroxyC1-C6 alkyl-. Nonlimitingexamples include 2-hydroxyethyl.

In one embodiment of Formula II, R³ is hydroxyC1-C6 alkoxy-. Nonlimitingexamples include 2-hydroxyethoxy and 2-hydroxypropoxy having thestructures:

-   -   respectively.

In one embodiment of Formula II, R³ is C1-C6 alkoxy. Nonlimitingexamples include methoxy, ethoxy, 1-methylethoxy, and2,2-dimethylethoxy.

In one embodiment of Formula II, R³ is fluoroC1-C6 alkoxy. A nonlimitingexample includes 2,2-difluoroethoxy.

In one embodiment of Formula II, R³ is C3-C6 cycloalkyl. A nonlimitingexample is cyclopropyl.

In one embodiment of Formula II, R³ is (C3-C6 cycloalkyl)C1-C6 alkoxy-.A nonlimiting example is cyclopropylmethoxy.

In one embodiment of Formula II, R³ is H or hydroxyC1-C6 alkoxy-.

In one embodiment of Formula II, R^(a) is halogen. In one embodiment ofFormula II, R^(a) is fluoro or chloro. In one embodiment of Formula II,R^(a) is fluoro.

In one embodiment of Formula II, R^(b) is fluoro, chloro, bromo, iodo,ethyl, propyl, isopropyl, methylthio, difluoromethylthio,trifluoromethyl, methoxy, difluoromethoxy, cyclopropyl, or CH₃C(═O)—.

In one embodiment of Formula II, R^(b) is halogen, C1-C6 alkyl, C1-C6alkylthio, or fluoroC1-C6 alkoxy.

In one embodiment of Formula II, R^(b) is bromo, iodo, ethyl,methylthio, or difluoromethoxy. In one embodiment of Formula II, R^(b)is methylthio.

In one embodiment of Formula II, R^(a) is fluoro and R^(b) ismethylthio.

In one embodiment of Formula II, R^(a) is halogen and R^(b) is halogen,C1-C6 alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl,C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, or C1-C6alkyl-C(═O)—.

In one embodiment of Formula II, R^(a) is halogen and R^(b) is halogen,C1-C6 alkyl, C1-C6 alkylthio, or fluoroC1-C6 alkoxy.

In one embodiment of Formula II, R^(a) is halogen and R^(b) is halogen.In one embodiment of Formula II, R^(a) is halogen, R^(b) is halogen, R¹is H and R² is H.

In one embodiment of Formula II, R^(a) is halogen and R^(b) is C1-C6alkyl. In one embodiment of Formula II, R^(a) is halogen, R^(b) is C1-C6alkyl, R¹ is H and R² is H.

In one embodiment of Formula II, R^(a) is halogen and R^(b) is C1-C6alkylthio. In one embodiment of Formula II, R^(a) is halogen, R^(b) isC1-C6 alkylthio, R¹ is H and R² is H.

In one embodiment of Formula II, R^(a) is halogen and R^(b) isfluoroC1-C6 alkylthio. In one embodiment of Formula II, R^(a) ishalogen, R^(b) is fluoroC1-C6 alkylthio, R¹ is H and R² is H.

In one embodiment of Formula II, R^(a) is fluoro, R^(b) is methylthio,R¹ is H and R² is H.

In one embodiment of Formula II, R^(a) is halogen and R^(b) isfluoroC1-C6 alkyl. In one embodiment of Formula II, R^(a) is halogen,R^(b) is fluoroC1-C6 alkyl, R¹ is H and R² is H.

In one embodiment of Formula II, R^(a) is halogen and R^(b) is C1-C6alkoxy. In one embodiment of Formula II, R^(a) is halogen, R^(b) isC1-C6 alkoxy, R¹ is H and R² is H.

In one embodiment of Formula II, R^(a) is halogen and R^(b) isfluoroC1-C6 alkoxy. In one embodiment of Formula II, R^(a) is halogen,R^(b) is fluoroC1-C6 alkoxy, R¹ is H and R² is H.

In one embodiment of Formula II, R^(a) is halogen and R^(b) is C3-C6cycloalkyl. In one embodiment of Formula II, R^(a) is halogen, R^(b) isC3-C6 cycloalkyl, R¹ is H and R² is H.

In one embodiment of Formula II, R^(a) is halogen and R^(b) is C1-C6alkyl-C(═O)—. In one embodiment of Formula II, R^(a) is halogen, R^(b)is C1-C6 alkyl-C(═O)—, R¹ is H and R² is H.

In one embodiment of Formula II, R¹ is H, R² is H or CH₃—, R³ is H orhydroxyC1-C6 alkoxy-, R^(a) is halogen and R^(b) is halogen, C1-C6alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, or C1-C6 alkyl-C(═O)—.

In one embodiment of Formula II, R¹ is H, R² is H or CH₃—, R³ is H orhydroxyC1-C6 alkoxy-, R^(a) is halogen, and R^(b) is halogen, C1-C6alkyl, C1-C6 alkylthio, or fluoroC1-C6 alkoxy.

In one embodiment of any of the above described embodiments of FormulaII, the group

is selected from the structures:

The term “compound,” as used herein is meant to include allstereoisomers, geometric isomers, tautomers, and isotopes of thestructures depicted. Compounds herein identified by name or structure asone particular tautomeric form are intended to include other tautomericforms unless otherwise specified.

The compounds of the formulae provided herein may have asymmetric carbonatoms. The carbon carbon bonds of the compounds of the invention may bedepicted herein using a solid line (

), a solid wedge (

), or a dotted wedge (

). The use of a solid line to depict bonds to asymmetric carbon atoms ismeant to indicate that all possible stereoisomers (e.g. specificenantiomers, racemic mixtures, etc.) at that carbon atom are included.The use of either a solid or dotted wedge to depict bonds to asymmetriccarbon atoms is meant to indicate that only the stereoisomer shown ismeant to be included. It is possible that compounds of the invention maycontain more than one asymmetric carbon atom. In those compounds, theuse of a solid line to depict bonds to asymmetric carbon atoms is meantto indicate that all possible stereoisomers are meant to be included andthe attached stereocenter. For example, unless stated otherwise, it isintended that the compounds of the invention can exist as enantiomersand diastereomers or as racemates and mixtures thereof. The use of asolid line to depict bonds to one or more asymmetric carbon atoms in acompound of the invention and the use of a solid or dotted wedge todepict bonds to other asymmetric carbon atoms in the same compound ismeant to indicate that a mixture of diastereomers is present.

Compounds of the invention that have chiral centers may exist asstereoisomers, such as racemates, enantiomers, or diastereomers.

Stereoisomers of the compounds of the formulae herein can include cisand trans isomers, optical isomers such as (R) and (S) enantiomers,diastereomers, geometric isomers, rotational isomers, atropisomers,conformational isomers, and tautomers of the compounds of the invention,including compounds exhibiting more than one type of isomerism; andmixtures thereof (such as racemates and diastereomeric pairs).

Also included are acid addition salts or base addition salts, whereinthe counterion is optically active, for example, d-lactate or l-lysine,or racemic, for example, dl-tartrate or dl-arginine.

When any racemate crystallizes, crystals of two different types arepossible. The first type is the racemic compound (true racemate)referred to above wherein one homogeneous form of crystal is producedcontaining both enantiomers in equimolar amounts. The second type is theracemic mixture or conglomerate wherein two forms of crystal areproduced in equimolar amounts each comprising a single enantiomer.

Conventional techniques for the preparation/isolation of individualenantiomers include chiral synthesis from a suitable optically pureprecursor or resolution of the racemate (or the racemate of a salt orderivative) using, for example, chiral high-pressure liquidchromatography (HPLC) or superfluid critical chromatography (SFC).

Alternatively, the racemate (or a racemic precursor) may be reacted witha suitable optically active compound, for example, an alcohol, or, inthe case where the compound contains an acidic or basic moiety, an acidor base such as tartaric acid or 1-phenylethylamine. The resultingdiastereomeric mixture may be separated by chromatography and/orfractional crystallization and one or both of the diastereoisomersconverted to the corresponding pure enantiomer(s) by means well known toone skilled in the art.

Chiral compounds of the invention (and chiral precursors thereof) may beobtained in enantiomerically enriched form using chromatography,typically HPLC, on an asymmetric resin with a mobile phase consisting ofa hydrocarbon, typically heptane or hexane, containing from 0 to 50%isopropanol, typically from 2 to 20%, and from 0 to 5% of an alkylamine,typically 0.1% diethylamine. Concentration of the eluent affords theenriched mixture.

Stereoisomeric conglomerates may be separated by conventional techniquesknown to those skilled in the art; see, for example, “Stereochemistry ofOrganic Compounds” by E L Eliel (Wiley, New York, 1994), the disclosureof which is incorporated herein by reference in its entirety.

The enantiomeric purity of compounds described herein may be describedin terms of enantiomeric excess (ee), which indicates the degree towhich a sample contains one enantiomer in greater amounts than theother. A racemic mixture has an ee of 0%, while a single completely pureenantiomer has an ee of 100%. Similarly, diastereomeric purity may bedescribed in terms of diastereomeric excess (de).

The compounds of the invention may exhibit the phenomena of tautomerismand structural isomerism. For example, the compounds may exist inseveral tautomeric forms, including the enol and imine form, and theketo and enamine form and geometric isomers and mixtures thereof. Allsuch tautomeric forms are included within the scope of compounds of theinvention. Tautomers exist as mixtures of a tautomeric set in solution.In solid form, usually one tautomer predominates. Even though onetautomer may be described, the present invention includes all tautomersof the compounds of the formulae provided. A tautomer of a compound ofFormula I may occur, for example, when R³ is hydrogen, that is:

In addition, some of the compounds of the invention may formatropisomers (e.g., substituted biaryls). Atropisomers areconformational stereoisomers which occur when rotation about a singlebond in the molecule is prevented, or greatly slowed, as a result ofsteric interactions with other parts of the molecule and thesubstituents at both ends of the single bond are unsymmetrical. Theinterconversion of atropisomers is slow enough to allow separation andisolation under predetermined conditions. The energy barrier to thermalracemization may be determined by the steric hindrance to free rotationof one or more bonds forming a chiral axis.

The present invention also includes pharmaceutically acceptableisotopically labeled compounds, which are identical to those recited inone of the formulae provided, but for the fact that one or more atomsare replaced by an atom having an atomic mass or mass number differentfrom the atomic mass or mass number usually found in nature.

Isotopically labeled compounds of the invention can generally beprepared by conventional techniques known to those skilled in the art orby processes analogous to those described herein, using an appropriateisotopically labeled reagent in place of the nonlabeled reagentotherwise employed.

Examples of isotopes that may be incorporated into compounds of theinvention include isotopes of hydrogen, carbon, nitrogen, oxygen,phosphorus, sulfur, fluorine and chlorine, such as, but not limited to,²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³²P, ³⁵S, ¹⁸F and ³⁶Cl. Certainisotopically labeled compounds of the invention, for example those intowhich radioactive isotopes such as ²H, ³H or ¹⁴C are incorporated, areuseful in one or both of drug or substrate tissue distribution assays.Tritiated, i.e., ³H, and carbon 14, i.e., ¹⁴C, isotopes are particularlypreferred for their ease of preparation and detectability. Further,substitution with heavier isotopes such as deuterium, i.e., 2H, canafford certain therapeutic advantages resulting from greater metabolicstability, for example increased in vivo half-life or reduced dosagerequirements and, hence, may be preferred in some circumstances.Substitution with positron emitting isotopes, such as ¹¹C, ¹⁸F, ¹⁵O and¹³N, may be useful in Positron Emission Topography (PET) studies forexamining substrate receptor occupancy. Isotopically labeled compoundsof the invention may generally be prepared by carrying out theprocedures disclosed in the Schemes and/or in the Examples andPreparations below, by substituting an isotopically labeled reagent fora non-isotopically labeled reagent.

Pharmaceutically acceptable solvates in accordance with the inventioninclude those wherein the solvent of crystallization may be isotopicallysubstituted, e.g., D₂O, d⁶-acetone, d⁶-DMSO.

Unless indicated otherwise, all references herein to the inventivecompounds include references to salts, solvates, hydrates and complexesthereof, and to solvates, hydrates and complexes of salts thereof,including polymorphs, stereoisomers, and isotopically labelled versionsthereof.

Compounds of the invention may exist in the form of pharmaceuticallyacceptable salts such as, e.g., acid addition salts and base additionsalts of the compounds of one of the formulae provided herein. As usedherein, the term “pharmaceutically acceptable salt” refers to thosesalts which retain the biological effectiveness and properties of theparent compound. The phrase “pharmaceutically acceptable salt(s)”, asused herein, unless otherwise indicated, includes salts of acidic orbasic groups which may be present in the compounds of the formulaedisclosed herein.

For example, the compounds of the invention that are basic in nature arecapable of forming a wide variety of salts with various inorganic andorganic acids. Although such salts must be pharmaceutically acceptablefor administration to animals, it is often desirable in practice toinitially isolate the compound of the present invention from thereaction mixture as a pharmaceutically unacceptable salt and then simplyconvert the latter back to the free base compound by treatment with analkaline reagent and subsequently convert the latter free base to apharmaceutically acceptable acid addition salt. The acid addition saltsof the base compounds of this invention can be prepared by treating thebase compound with a substantially equivalent amount of the selectedmineral or organic acid in an aqueous solvent medium or in a suitableorganic solvent, such as methanol or ethanol. Upon evaporation of thesolvent, the desired solid salt is obtained. The desired acid salt canalso be precipitated from a solution of the free base in an organicsolvent by adding an appropriate mineral or organic acid to thesolution.

The acids that may be used to prepare pharmaceutically acceptable acidaddition salts of such basic compounds of those that form nontoxic acidaddition salts, i.e., salts containing pharmacologically acceptableanions, such as the hydrochloride, hydrobromide, hydroiodide, nitrate,sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate,lactate, salicylate, citrate, acid citrate, tartrate, pantothenate,bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate,gluconate, glucuronate, saccharate, formate, benzoate, glutamate,methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonateand pamoate salts.

Examples of salts include, but are not limited to, acetate, acrylate,benzenesulfonate, benzoate (such as chlorobenzoate, methylbenzoate,dinitrobenzoate, hydroxybenzoate, and methoxybenzoate), bicarbonate,bisulfate, bisulfite, bitartrate, borate, bromide, butyne-1,4-dioate,calcium edetate, camsylate, carbonate, chloride, caproate, caprylate,clavulanate, citrate, decanoate, dihydrochloride, dihydrogenphosphate,edetate, edislyate, estolate, esylate, ethylsuccinate, formate,fumarate, gluceptate, gluconate, glutamate, glycollate,glycollylarsanilate, heptanoate, hexyne-1,6-dioate, hexylresorcinate,hydrabamine, hydrobromide, hydrochloride, g hydroxy butyrate, iodide,isobutyrate, isothionate, lactate, lactobionate, laurate, malate,maleate, malonate, mandelate, mesylate, metaphosphate, methanesulfonate, methylsulfate, monohydrogenphosphate, mucate, napsylate,naphthalene-1-sulfonate, naphthalene-2-sulfonate, nitrate, oleate,oxalate, pamoate (embonate), palmitate, pantothenate, phenylacetates,phenylbutyrate, phenylpropionate, phthalate, phosphate/diphosphate,polygalacturonate, propanesulfonate, propionate, propiolate,pyrophosphate, pyrosulfate, salicylate, stearate, subacetate, suberate,succinate, sulfate, sulfonate, sulfite, tannate, tartrate, teoclate,tosylate and valerate salts.

Illustrative examples of suitable salts include organic salts derivedfrom amino acids, such as glycine and arginine, ammonia, primary,secondary, and tertiary amines and cyclic amines, such as piperidine,morpholine and piperazine, and inorganic salts derived from sodium,calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminumand lithium.

The compounds of the invention that include a basic moiety, such as anamino group, may form pharmaceutically acceptable salts with variousamino acids, in addition to the acids mentioned above.

Alternatively, the compounds useful that are acidic in nature may becapable of forming base salts with various pharmacologically acceptablecations. Examples of such salts include the alkali metal or alkalineearth metal salts and particularly, the sodium and potassium salts.These salts are all prepared by conventional techniques. The chemicalbases which are used as reagents to prepare the pharmaceuticallyacceptable base salts of this invention are those which form nontoxicbase salts with the acidic compounds herein. These salts may be preparedby any suitable method, for example, treatment of the free acid with aninorganic or organic base, such as an amine (primary, secondary ortertiary), an alkali metal hydroxide or alkaline earth metal hydroxide,or the like. These salts can also be prepared by treating thecorresponding acidic compounds with an aqueous solution containing thedesired pharmacologically acceptable cations, and then evaporating theresulting solution to dryness, preferably under reduced pressure.Alternatively, they may also be prepared by mixing lower alkanolicsolutions of the acidic compounds and the desired alkali metal alkoxidetogether, and then evaporating the resulting solution to dryness in thesame manner as before. In either case, stoichiometric quantities ofreagents are preferably employed in order to ensure completeness ofreaction and maximum yields of the desired final product.

The chemical bases that may be used as reagents to preparepharmaceutically acceptable base salts of the compounds of the inventionthat are acidic in nature are those that form nontoxic base salts withsuch compounds. Such nontoxic base salts include, but are not limitedto, those derived from such pharmacologically acceptable cations such asalkali metal cations (e.g., potassium and sodium) and alkaline earthmetal cations (e.g., calcium and magnesium), ammonium or water solubleamine addition salts such as N-methylglucamine (meglumine), and thelower alkanolammonium and other base salts of pharmaceuticallyacceptable organic amines.

Hemisalts of acids and bases may also be formed, for example,hemisulphate and hemicalcium salts.

For a review on suitable salts, see Handbook of Pharmaceutical Salts:Properties, Selection, and Use by Stahl and Wermuth (Wiley VCH, 2002).Methods for making pharmaceutically acceptable salts of compounds of theinvention, and of interconverting salt and free base forms, are known toone of skill in the art.

Salts of the present invention can be prepared according to methodsknown to those of skill in the art. A pharmaceutically acceptable saltof the inventive compounds can be readily prepared by mixing togethersolutions of the compound and the desired acid or base, as appropriate.The salt may precipitate from solution and be collected by filtration ormay be recovered by evaporation of the solvent. The degree of ionizationin the salt may vary from completely ionized to almost nonionized.

It will be understood by those of skill in the art that the compounds ofthe invention in free base form having a basic functionality may beconverted to the acid addition salts by treating with a stoichiometricexcess of the appropriate acid. The acid addition salts of the compoundsof the invention may be reconverted to the corresponding free base bytreating with a stoichiometric excess of a suitable base, such aspotassium carbonate or sodium hydroxide, typically in the presence ofaqueous solvent, and at a temperature of between about 0° C. and 100° C.The free base form may be isolated by conventional means, such asextraction with an organic solvent. In addition, acid addition salts ofthe compounds of the invention may be interchanged by taking advantageof differential solubilities of the salts, volatilities or acidities ofthe acids, or by treating with the appropriately loaded ion exchangeresin. For example, the interchange may be affected by the reaction of asalt of the compounds of the invention with a slight stoichiometricexcess of an acid of a lower pK than the acid component of the startingsalt. This conversion is typically carried out at a temperature betweenabout 0° C. and the boiling point of the solvent being used as themedium for the procedure. Similar exchanges are possible with baseaddition salts, typically via the intermediacy of the free base form.

The compounds of the invention may exist in both unsolvated and solvatedforms. When the solvent or water is tightly bound, the complex will havea well-defined stoichiometry independent of humidity. When, however, thesolvent or water is weakly bound, as in channel solvates and hygroscopiccompounds, the water/solvent content will be dependent on humidity anddrying conditions. In such cases, non-stoichiometry will be the norm.The term ‘solvate’ is used herein to describe a molecular complexcomprising the compound of the invention and one or morepharmaceutically acceptable solvent molecules, for example, ethanol. Theterm ‘hydrate’ is employed when the solvent is water. Pharmaceuticallyacceptable solvates in accordance with the invention include hydratesand solvates wherein the solvent of crystallization may be isotopicallysubstituted, e.g. D₂O, d6-acetone, d6-DMSO.

The invention also relates to prodrugs of the compounds of the formulaeprovided herein. Thus, certain derivatives of compounds of the inventionwhich may have little or no pharmacological activity themselves can,when administered to a patient, be converted into the inventivecompounds, for example, by hydrolytic cleavage. Such derivatives arereferred to as ‘prodrugs’. Further information on the use of prodrugsmay be found in ‘Prodrugs as Novel Delivery Systems, Vol. 14, ACSSymposium Series (T Higuchi and W Stella); ‘Bioreversible Carriers inDrug Design’, Pergamon Press, 1987 (ed. E B Roche, AmericanPharmaceutical Association); Guarino, V. R; Stella, V. J.: BiotechPharm. Aspects 2007 5 (Pt2) 133-187; and J. Rautio et al. Nature ReviewsDrug Discovery, 17, 559-587 (2018) the disclosures of which areincorporated herein by reference in their entireties.

Prodrugs in accordance with the invention can, for example, be producedby replacing appropriate functionalities present in the inventivecompounds with certain moieties known to those skilled in the art as‘promoieties’ as described, for example, in “Design of Prodrugs” by HBundgaard (Elsevier, 1985), the disclosure of which is incorporatedherein by reference in its entirety.

Some non-limiting examples of prodrugs in accordance with the inventioninclude:

-   -   (i) when a compound contains a carboxylic acid functionality        (—COOH), an ester thereof, for example, replacement of the        hydrogen with (C1-C6)alkyl;    -   (ii) when a compound contains an alcohol functionality (—OH), an        ether thereof, for example, replacement of the hydrogen with        (C1-C6)alkanoyloxymethyl, or with a phosphate ether group; and    -   (iii) when a compound contains a primary or secondary amino        functionality (NH₂ or NHR where R is not H), an amide thereof,        for example, replacement of one or both hydrogens with a        suitably metabolically labile group, such as an amide,        carbamate, urea, phosphonate, sulfonate, etc.

Further examples of replacement groups in accordance with the foregoingexamples and examples of other prodrug types may be found in theaforementioned references.

Finally, certain inventive compounds may themselves act as prodrugs ofother of the inventive compounds.

Also included within the scope of the invention are metabolites ofcompounds of the formulae described herein, i.e., compounds formed invivo upon administration of the drug, often by oxidation ordealkylation. Some examples of metabolites in accordance with theinvention include, but are not limited to,

-   -   (i) where the compound of the invention contains an alkyl group,        a hydroxyalkyl derivative thereof (—CH>—COH):    -   (ii) where the compound of the invention contains an alkoxy        group, a hydroxy derivative thereof (—OR->—OH);    -   (iii) where the compound of the invention contains a tertiary        amino group, a secondary amino derivative thereof (—NRR′->—NHR        or —NHR′);    -   (iv) where the compound of the invention contains a secondary        amino group, a primary derivative thereof (—NHR->—NH₂);    -   (v) where the compound of the invention contains a phenyl        moiety, a phenol derivative thereof (-Ph->-PhOH);    -   (vi) where the compound of the invention contains an amide        group, a carboxylic acid derivative thereof (—CONH₂->COOH); and    -   (vii) where the compound contains a hydroxy or carboxylic acid        group, the compound may be metabolized by conjugation, for        example with glucuronic acid to form a glucuronide. Other routes        of conjugative metabolism exist. These pathways are frequently        known as Phase 2 metabolism and include, for example, sulfation        or acetylation. Other functional groups, such as NH groups, may        also be subject to conjugation.

In one embodiment, provided herein are solid forms of8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.In one embodiment, the solid form is a crystalline form. In oneembodiment, the solid form is an amorphous form.

The terms “crystalline” as used herein, means having a regularlyrepeating arrangement of molecules or external face planes. A singlecompound may give rise to a variety of crystalline forms where each formhas different and distinct solid state physical properties, such asdifferent solubility profiles, dissolution rates, melting pointtemperatures, flowability, and/or different X-ray diffraction peaks. Thedifferences in physical properties may affect pharmaceutical parameterssuch as storage stability, compressibility and density (which can beimportant in formulation and product manufacturing), and dissolutionrate (which can be an important factor in bioavailability).

The term ‘amorphous’ refers to a state in which the material lacks longrange order at the molecular level and, depending upon temperature, mayexhibit the physical properties of a solid or a liquid. Typically, suchmaterials do not give distinctive X-ray diffraction patterns and, whileexhibiting the properties of a solid, are more formally described as aliquid. Upon heating, a change from solid to liquid properties occurswhich is characterized by a change of state, typically second order(‘glass transition’).

There are a number of analytical methods one of ordinary skill in theart in solid-state chemistry can use to analyze solid forms. PowderX-ray diffraction may also be suitable for quantifying the amount of acrystalline solid form (or forms) in a mixture. In powder X-raydiffraction, X-rays are directed onto a crystalline powder and theintensity of the diffracted X-rays is measured as a function of theangle between the X-ray source and the beam diffracted by the sample.The intensity of these diffracted X-rays can be plotted on a graph aspeaks with the x-axis being the angle (this is known as the “2-theta”angle) between the X-ray source and the diffracted X-rays and with they-axis being the intensity of the diffracted X-rays. This graph iscalled a powder X-ray diffraction pattern or powder pattern. Differentcrystalline solid forms exhibit different powder patterns because thelocation of the peaks on the x-axis is a property of the solid-statestructure of the crystal.

One of ordinary skill in the art will appreciate that a typicalprecision of the 2-theta x-axis value of a peak in a powder pattern ison the order of plus or minus 0.2 degrees 2-theta (±0.2 degrees 2theta). Thus, for example, a diffraction peak that appears at “about18.0 degrees 2-theta” means that the peak appears at 18.0 degrees±0.2degrees 2-theta, i.e., it could be between 17.8 degrees 2-theta and 18.2degrees 2-theta when measured on most X-ray diffractometers under mostconditions. Further, one skilled in the art will appreciate that therelative peak intensities will show inter-apparatus variability as wellas variability due to degree of crystallinity, preferred orientation,prepared sample surface, and other factors known to those skilled in theart and should be taken as qualitative measures only. Accordingly, asused herein, the term “essentially the same” with reference to powderX-ray diffraction peak positions, means that typical variability in peakposition and intensity are on the order of ±0.2 degrees 2 theta.

Powder X-ray diffraction is just one of several analytical techniquesone may use to characterize and/or identify crystalline solid forms.Spectroscopic techniques such as Raman (including microscopic Raman),infrared, and solid state NMR spectroscopies may be used to characterizeand/or identify crystalline solid forms. These techniques may also beused to quantify the amount of one or more crystalline solid forms in amixture and peak values can also be reported with the modifier “about”in front of the peak values.

The term “anhydrous” as used herein, refers to a crystalline formwithout any solvent or water molecules in the crystal lattice

The term “hydrate” refers to a solvate comprising the compound and astoichiometric or non-stoichiometric amount of water. The term“monohydrate” refers to a hydrate comprising one molecule of water permolecule of compound (i.e., a 1:1 stoichiometry of water to compound).

In one embodiment, the invention provides crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 1.

In one embodiment, crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 1 is characterized by powder X-ray diffraction (PXRD) (2-theta).

Table X provides a PXRD peak list for crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 1 in degrees 2-theta (±0.2 degrees 2-theta).

TABLE X Angle Rel. Intensity (degrees 2-Theta) (%)  5.0 8.1  8.7 7.5 9.3 18.3 10.8 100.0 14.5 8.1 15.3 17.4 18.8 9.9 20.5 8.4

In one embodiment, the invention provides crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 1 having a PXRD pattern comprising characterizing peaks at 5.0,8.7, 9.3, 10.8, 14.5, 15.3, 18.8 and 20.5 degrees 2-theta (±0.2 degrees2-theta).

In one embodiment, the invention provides crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 1 having a PXRD pattern comprising peaks at 2-theta valuesessentially the same as shown in FIG. 1 .

In one embodiment, the invention provides crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2.

In one embodiment, crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2 is characterized by powder X-ray diffraction (PXRD) (2-theta). Inone embodiment, PXRD analysis of crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2 is conducted at 25° C. and at a relative humidity below 10%, forexample as described in Example 77.

Table Y provides a PXRD peak list for crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2 in degrees 2-theta (±0.2 degrees 2-theta).

TABLE Y Angle Rel. Intensity (degrees 2-Theta) (%)  7.1 5.2  9.4 100.012.4 8.8 12.8 5.1 14.3 14.2 15.6 14.8 16.4 3.9 17.4 5.3 18.5 7.3 18.943.4 19.5 17.7 19.9 35.2 21.1 20.7 21.4 7.9 23.2 12.1 23.7 9.1 24.8 6.225.6 31.2 27.6 8.5 30.3 4.6 33.2 5.5 33.5 5.3 37.5 6.6

In one embodiment, the invention provides crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2 having a PXRD pattern comprising characterizing peaks at 7.1,9.4, 12.4, 12.8, 14.3, 15.6, 16.4, 17.4, 18.5, 18.9, 19.5, 19.9, 21.1,21.4, 23.2, 23.7, 24.8, 25.6, 27.6, 30.3, 33.2, 33.5, and 37.5 degrees2-theta (±0.2 degrees 2-theta).

In one embodiment, the invention provides crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2 having a PXRD pattern comprising peaks at 2-theta valuesessentially the same as shown in FIG. 2 .

In one embodiment, the invention provides crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3.

In one embodiment, crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3 is characterized by powder X-ray diffraction (PXRD) (2-theta). Inone embodiment, PXRD analysis of crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3 is conducted at 25° C. and at relative humidity above 30%.

Table Z provides a PXRD peak list for crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3 in degrees 2-theta (±0.2 degrees 2-theta).

TABLE Z Angle Rel. Intensity (degrees 2-Theta) (%)  6.9 10.0  9.1 100.011.8 6.0 12.0 21.1 13.7 11.6 14.0 18.1 15.2 31.2 15.8 4.3 18.0 72.2 18.319.3 19.0 27.7 19.3 7.6 20.2 37.2 20.9 8.8 21.6 49.2 22.6 19.2 23.6 30.524.0 27.9 24.9 18.1 25.2 12.3 25.8 69.7 27.5 32.0 28.1 11.5 28.4 5.929.8 15.0 30.9 11.2 31.7 6.2 32.3 7.3 36.5 13.2

In one embodiment, the invention provides crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3 having a PXRD pattern comprising characterizing peaks at 13.7,18.0 and 18.3 degrees 2-theta (±0.2 degrees 2-theta).

In one embodiment, the invention provides crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3 having a PXRD pattern comprising characterizing peaks at 6.9,9.1, 13.7, 18.0 and 18.3 degrees 2-theta (±0.2 degrees 2-theta).

In one embodiment, the invention provides crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3 having a PXRD pattern comprising characterizing peaks at 6.9,9.1, 11.8, 12.0, 13.7, 14.0, 15.2, 15.8, 18.0, 18.3, 19.0, 19.3, 20.2,20.9, 21.6, 22.6, 23.6, 24.0, 24.9, 25.2, 25.8, 27.5, 28.1, 28.4, 29.8,30.9, 31.7, 32.3 and 36.5 degrees 2-theta (±0.2 degrees 2-theta).

In one embodiment, the invention provides crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3 having a PXRD pattern comprising peaks at 2-theta valuesessentially the same as shown in FIG. 3 .

In one embodiment, the invention provides amorphous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 4.

In one embodiment, amorphous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 4 is characterized by powder X-ray diffraction (PXRD) (2-theta).

In one embodiment, the invention provides amorphous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 4 having a PXRD pattern comprising peaks at 2-theta valuesessentially the same as shown in FIG. 4 .

The invention further provides therapeutic methods and uses comprisingadministering the compounds of the invention, or pharmaceuticallyacceptable salts thereof, alone or in combination with other therapeuticagents or palliative agents.

Compounds of Formula I and Formula II, and pharmaceutically acceptablesalts thereof are useful for treating diseases and disorders which canbe treated with a MEK kinase inhibitor, such as MEK-associated diseasesand disorders, e.g., for the treatment of abnormal cell growth, suchtumors, for example MEK-associated tumors. The ability of compounds ofFormula I and Formula II, and pharmaceutically acceptable salts thereof,to act as MEK inhibitors may be demonstrated by the assay described inExample A. IC₅₀ values are shown in Table A.

Accordingly, in one embodiment, provided herein is a method of treatinga tumor, comprising administering to a subject in need thereof atherapeutically effective amount of a compound of Formula I or apharmaceutically acceptable salt thereof, or a compound of Formula II ora pharmaceutically acceptable salt thereof. In one embodiment, the tumoris a MEK-associated tumor.

As used herein, the terms “MEK kinase inhibitor” and “MEK inhibitor” areused interchangeably and refer to a compound that inhibits themitogen-activated protein kinase kinase enzymes MEK1 and/or MEK2.

The terms “MEK-associated” and “MEK-mediated” are used interchangeablyand refer to a disease or disorder having constitutive activation of aMEK kinase which can be treated with a MEK inhibitor. Examples include aMEK-associated abnormal cell growth such as MEK-associated tumors, e.g.,MEK-associated cancers. In one embodiment, the term “MEK-associated”refers to a disease or disorder having a dysregulation of the expressionor activity of a MEK kinase, or a dysregulation of a BRAF gene or a BRAFkinase.

The phrase “dysregulation of the expression or activity of a MEK kinase”refers to gene amplification that results in overexpression of a MEKprotein or an autocrine activity resulting from the overexpression of aMEK gene in a cell that results in a pathogenic increase in the activityof a kinase domain of a MEK protein (e.g., a constitutively activekinase domain of a MEK protein) in a cell.

The phrase “dysregulation of a BRAF gene or a BRAF kinase” refers to agenetic mutation (e.g., a BRAF gene translocation that results in theexpression of a fusion protein, a deletion in a BRAF gene that resultsin the expression of a BRAF protein that includes a deletion of at leastone amino acid as compared to the wild-type BRAF protein, or a mutationin a BRAF gene that results in the expression of a BRAF protein with oneor more point mutations as compared to a wild-type BRAF protein). Asanother example, a dysregulation of a BRAF gene, a BRAF protein, orexpression or activity, or level of any of the same, can be a mutationin a BRAF gene that encodes a BRAF protein that is constitutively activeor has increased activity as compared to a protein encoded by a BRAFgene that does not include the mutation. For example, a dysregulation ofa BRAF gene, a BRAF protein, or expression or activity, or level of anyof the same, can be the result of a gene or chromosome translocationwhich results in the expression of a fusion protein that contains afirst portion of BRAF that includes a functional kinase domain, and asecond portion of a partner protein (i.e., that is not BRAF).

In one embodiment, the MEK-associated disease or disorder has anactivating BRAF mutation. In one embodiment, the MEK-associated diseaseor disorder is a MEK-associated cancer having an activating BRAFmutation. Non-limiting examples of BRAF mutations include BRAF V600mutations, e.g., V600E, V600D, V600K, V600R and V600S. In oneembodiment, the BRAF mutation is a V600E mutation. In one embodiment,the BRAF mutation is a V600K mutation.

In one embodiment, the MEK-associated disease or disorder is aMEK-associated tumor has one or more BRAF fusions that lead toconstitutive kinase activation and transformation, including but notlimited to KIAA11549-BRAF, MKRN1-BRAF, TRIM24-BRAF, AGAP3-BRAF,ZC3HAV1-BRAF, AKAP9-BRAF, CCDC6-BRAF, AGK-BRAF, EPS15-BRAF, NUP214-BRAF,ARMC10-BRAF, BTF3L4-BRAF, GHR-BRAF, ZC3HAV1-BRAF, ZNF767-BRAF,CCDC91-BRAF, DYNC112-BRAF, ZKSCAN1-BRAF, GTF21-BRAF, MZT1-BRAF,RAD18-BRAF, CUX1-BRAF, SLC12A7-BRAF, MYRIP-BRAF, SND1-BRAF, NUB1-BRAF,KLHL7-BRAF, TANK-BRAF, RBMS3-BRAF, STRN3-BRAF, STK35-BRAF, ETFA-BRAF,SVOPL-BRAF, JHDM1D-BRAF, or BCAP29-BRAF.

In one embodiment, the MEK-associated disease or disorder is aMEK-associated tumor having a BRAF-fusion protein, wherein the tumor isbreast carcinoma (e.g., breast invasive ductal carcinoma) colorectalcarcinoma (e.g., colon adenocarcinoma), esophageal carcinoma (e.g.,esophagus adenocarcinoma), glioma (e.g., brain desmoplastic infantileganglioglioma, brain pilocytic astrocytoma, brain pleomorphicxanthoastrocytoma, spinal cord low-grade glioma (NOS), anaplasticoligodendroglioma, anaplastic ganglioglioma), head & neck carcinoma(e.g., head and neck neuroendocrine carcinoma), lung carcinoma (e.g.,lung adenocarcinoma, lung non-small-cell lung cancer (NOS)), melanoma(e.g., cutaneous melanoma Spitzoid, mucosal melanoma non-Spitzoid,cutaneous melanoma Spitzoid, unknown primary melanoma, cutaneousmelanoma non-Spitzoid), pancreatic carcinoma (e.g., adenocarcinoma,pancreas acinar cell carcinoma), prostatic carcinoma (e.g., prostateacinar adenocarcinoma), sarcoma (malignant solid fibrous tumor), thyroidcarcinoma (thyroid papillary carcinoma), unknown primary carcinoma(e.g., unknown primary, adenocarcinoma), pleura mesothelioma, rectumadenocarcinoma, uterus endometrial carcinoma (e.g., uterus endometrialadenocarcinoma (NOS)) or ovary serous carcinoma.

In one embodiment, the MEK-associated cancer is selected from thecancers having the BRAF-fusion proteins described in Table 1 (J. S.Ross, et al., Int. J. Cancer: 138, 881-890 (2016)).

TABLE 1 Exemplary BRAF Fusion Partners and Cancers Tumor group histologytumor type fusion breast carcinoma BCAP29-BRAF breast carcinoma breastcarcinoma metastatic KIAA11549-BRAF colorectal carcinoma colonadenocarcinoma primary MKRN1-BRAF colorectal carcinoma colonadenocarcinoma metastatic TRIM24-BRAF colorectal carcinoma coionadenocarcinoma primary AGAP3-BRAF esophageal esophagus adenocarcinomaprimary ZC3HAV1-BRAF carcinoma glioma brain desmoplastic infantileprimary KIAA11549-BRAF ganglioglioma glioma brain pilocytic astrocytomaprimary KIAA11549-BRAF glioma brain pleomorphic primary KIAA11549-BRAFxanthoastrocytoma glioma spinal cord low-grade glioma primaryKIAA11549-BRAF (NOS) glioma brain pilocytic astrocytoma primaryAKAP9-BRAF glioma brain pleomorphic primary CCDC6-BRAF xanthoastrocytomaglioma brain pleomorphic primary AGK-BRAF xanthoastrocytoma glioma notpilocytic; anaplastic primary AGK-BRAF oligodendroglioma glioma notpilocytic; anaplastic primary KIAA11549-BRAF ganglioglioma head & neckhead and neck primary MKRN1-BRAF carcinoma neuroendocrine carcinoma lungcarcinoma lung adenocarcinoma metastatic EPS15-BRAF lung carcinoma lungnon-small cell lung primary NUP214-BRAF cancer (NOS) lung carcinoma lungadenocarcinoma primary ARMC10-BRAF lung carcinoma lung adenocarcinomaprimary BTF3L4-BRAF lung carcinoma lung adenocarcinoma primary AGK-BRAFlung carcinoma lung adenocarcinoma metastatic GHR-BRAF lung carcinomalung adenocarcinoma primary ZC3HAV1-BRAF lung carcinoma lung non-smallcell lung primary TRIM224-BRAF cancer (NOS) melanoma cutaneous melanomaSpitzoid primary TRIM24-BRAF melanoma mucosal melanoma non- metastaticZNF767-BRAF Spitzoid melanoma cutaneious melanoma non- metastaticCCDC91-BRAF Spitzoid melanoma cutaneous melanoma Spitzoid primaryDYNC112-BRAF melanoma cutaneous melanoma Spitzoid metastatic AKAP9-BRAFmelanoma cutaneous melanoma Spitzoid metastatic ZKSCAN1-BRAF melanomaunknown primary melanoma metastatic GTF2I-BRAF melanoma cutaneousmelanoma non- metastatic AGAP3-BRAF Spitzoid melanoma cutaneous melanomaSpitzoid metastatic AGK-BRAF melanoma cutaneous melanoma Spitzoidmetastatic MZT1-BRAF melanoma cutaneious melanoma non- primaryRAD18-BRAF Spitzoid melanoma cutaneous melanoma Spitzoid metastaticCUX1-BRAF melanoma cutaneous melanoma Spitzoid metastatic SLC12A7-BRAFpancreatic pancreas ductal primary MYRIP-BRAF carcinoma adenocarcinomapancreatic pancreas acinar cell metastatic SND1-BRAF carcinoma carcinomaprostatic carcinoma prostate acinar metastatic NUB1-BRAF adenocarcinomasarcoma malignant solid fibrous tumor primary KIAA1549-BRAF thyroidcarcinoma thyroid papillary carcinoma primary KLHL7-BRAF thyroidcarcinoma thyroid papillary carcinoma primary TANK-BRAF thyroidcarcinoma thyroid papillary carcinoma metastatic RBMS3-BRAF unknownprimary unknown primary, metastatic STRN3-BRAF carcinoma adenocarcinomaunknown primary unknown primary, carcinoma metastatic SND1-BRAFcarcinoma (NOS) pleura pleura mesothelioma primary STK35-BRAFmesothelioma rectum rectum adenocarcinoma metastatic ETFA-BRAFadenocarcinoma uterus endometrial uterus endometrial metastaticSVOPL-BRAF carcinoma adenocarcinoma (NOS) ovary serous ovary serouscarcinoma metastatic JHDM1D-BRAF carcinoma

In one embodiment, the MEK-associated tumor is a BRAF wild-type tumor.

The term “wild-type” describes a nucleic acid (e.g., a BRAF gene or aBRAF mRNA) that is typically found in a subject that does not have adisease or disorder related to the reference nucleic acid or protein.

The term “wild-type BRAF” describes a BRAF nucleic acid (e.g., a BRAFgene or a BRAF mRNA) or a BRAF protein that is found in a subject thatdoes not have an activating BRAF-mutation.

“Abnormal cell growth”, as used herein, unless otherwise indicated,refers to cell growth that is independent of normal regulatorymechanisms (e.g., loss of contact inhibition), for example, a tumor.Abnormal cell growth may be benign (not cancerous), or malignant(cancerous).

The terms “cancer” or “cancerous” refers to any malignant and/orinvasive growth or tumor caused by abnormal cell growth. Cancer includesprimary cancer that originates at a specific site in the body, ametastatic cancer that has spread from the place in which it started toother parts of the body, a recurrence from the original primary cancerafter remission, and a second primary cancer that is a new primarycancer in a patient with a history of previous cancer of a differenttype from the second primary cancer. Cancer includes solid tumors namedfor the type of cells that form them, cancer of blood, bone marrow, orthe lymphatic system. A solid tumor is an abnormal growth or mass oftissue that usually does not contain cysts or liquid areas. Examples ofsolid tumors are sarcomas, carcinomas, and lymphomas. Leukemias (cancersof the blood) generally do not form solid tumors (National CancerInstitute, Dictionary of Cancer Terms).

As used herein, terms “treat” or “treatment” refer to therapeutic orpalliative measures. Beneficial or desired clinical results include, butare not limited to, alleviation, in whole or in part, of symptomsassociated with a disease or disorder or condition, diminishment of theextent of disease, stabilized (i.e., not worsening) state of disease,delay or slowing of disease progression, amelioration or palliation ofthe disease state (e.g., one or more symptoms of the disease), andremission (whether partial or total), whether detectable orundetectable.

The term “treat” or “treating” a cancer as used herein means toadminister a compound of the present invention to a subject havingcancer, or diagnosed with cancer, to achieve at least one positivetherapeutic effect, such as, for example, reduced number of cancercells, reduced tumor size, reduced rate of cancer cell infiltration intoperipheral organs, or reduced rate of tumor metastases or tumor growth,reversing, alleviating, or inhibiting the progress of, the disorder orcondition to which such term applies, or one or more symptoms of suchdisorder or condition. The term “treatment”, as used herein, unlessotherwise indicated, refers to the act of treating as “treating” isdefined immediately above. The term “treating” also includes adjuvantand neo-adjuvant treatment of a subject.

For the purposes of this invention, beneficial or desired clinicalresults include, but are not limited to, one or more of the following:reducing the proliferation of (or destroying) neoplastic or cancerouscell; inhibiting metastasis or neoplastic cells; shrinking or decreasingthe size of a tumor; an increase in the period of remission for asubject (e.g., as compared to the one or more metric(s) in a subjecthaving a similar cancer receiving no treatment or a different treatment,or as compared to the one or more metric(s) in the same subject prior totreatment); decreasing symptoms resulting from the cancer; increasingthe quality of life of those suffering from the cancer; decreasing thedose of other medications required to treat the cancer; delaying theprogression of the cancer; curing the cancer; overcoming one or moreresistance mechanisms of the cancer; and/or prolonging survival ofpatients the cancer. Positive therapeutic effects in cancer can bemeasured in several ways (see, for example, W. A. Weber, Assessing tumorresponse to therapy, J. Nucl. Med. 50 Suppl. 1:1 S-10S (2009). Forexample, with respect to tumor growth inhibition (T/C), according to theNational Cancer Institute (NCI) standards, a T/C less than or equal to42% is the minimum level of anti-tumor activity. A T/C<10% is considereda high anti-tumor activity level, with T/C (%)=median tumor volume ofthe treated/median tumor volume of the control×100.

In one embodiment, the treatment achieved by administration of acompound of the invention is defined by reference to any of thefollowing: partial response (PR), complete response (CR), overallresponse (OR), progression free survival (PFS), disease free survival(DFS) and overall survival (OS). PFS, also referred to as “Time to TumorProgression” indicates the length of time during and after treatmentthat the cancer does not grow and includes the amount of time patientshave experienced a CR or PR, as well as the amount of time patients haveexperienced stable disease (SD). DFS refers to the length of time duringand after treatment that the patient remains free of disease. OS refersto a prolongation in life expectancy as compared to naive or untreatedsubjects or patients. In one embodiment, response to treatment with acompound of the invention is any of PR, CR, OR, PFS, DFS, or OS that isassessed using Response Evaluation Criteria in Solid Tumors (RECIST) 1.1response criteria.

The treatment regimen for a compound of the invention that is effectiveto treat a cancer patient may vary according to factors such as thedisease state, age, and weight of the patient, and the ability of thetherapy to elicit an anti-cancer response in the subject. While anembodiment of any of the aspects of the invention may not be effectivein achieving a positive therapeutic effect in every subject, it shoulddo so in a statistically significant number of subjects as determined byany statistical test known in the art such as the Student's t-test, thechi2-test the U-test according to Mann and Whitney, the Kruskal-Wallistest (H-test), Jonckheere-Terpstrat-testy and the Wilcon on-test.

The terms “treatment regimen”, “dosing protocol” and “dosing regimen”are used interchangeably to refer to the dose and timing ofadministration of a compound of the invention, alone or in combinationwith another therapeutic agent.

“Ameliorating” means a lessening or improvement of one or more symptomsupon treatment with a combination described herein, as compared to notadministering the combination. “Ameliorating” also includes shorteningor reduction in duration of a symptom.

As used herein, the term “subject” refers to any animal, includingmammals such as humans. In one embodiment, the subject has experiencedand/or exhibited at least one symptom of the disease or disorder to betreated and/or prevented. In one embodiment, the subject has beenidentified or diagnosed as having a MEK-associated tumor (e.g., asdetermined using a regulatory agency-approved, e.g., FDA-approved, assayor kit). In one embodiment, the subject has a MEK-associated tumor thatis positive for a BRAF mutation (e.g., as determined using a regulatoryagency-approved assay or kit). The subject can be a subject whose tumorshave a MEK mutation (e.g., where the tumor is identified as such using aregulatory agency-approved, e.g., FDA-approved, kit or assay). In oneembodiment, the subject is suspected of having a MEK-associated tumor.In one embodiment, the subject has a clinical record indicating that thesubject has a MEK-associated tumor that has a BRAF mutation (andoptionally the clinical record indicates that the subject should betreated with any of the compositions provided herein). In oneembodiment, the subject is a human. In one embodiment, the human subjectis a pediatric subject.

The term “pediatric subject” as used herein refers to a subject underthe age of 21 years at the time of diagnosis or treatment. The term“pediatric” can be further be divided into various subpopulationsincluding: neonates (from birth through the first month of life);infants (1 month up to two years of age); children (two years of age upto 12 years of age); and adolescents (12 years of age through 21 yearsof age (up to, but not including, the twenty-second birthday)). BerhmanR E, Kliegman R, Arvin A M, Nelson W E, Nelson Textbook of Pediatrics,15th Ed. Philadelphia: W.B. Saunders Company, 1996; Rudolph A M, et al.Rudolph's Pediatrics, 21st Ed. New York: McGraw-Hill, 2002; and Avery MD, First L R. Pediatric Medicine, 2nd Ed. Baltimore: Williams & Wilkins;1994. In one embodiment, a pediatric subject is from birth through thefirst 28 days of life, from 29 days of age to less than two years ofage, from two years of age to less than 12 years of age, or 12 years ofage through 21 years of age (up to, but not including, the twenty-secondbirthday). In one embodiment, a pediatric subject is from birth throughthe first 28 days of life, from 29 days of age to less than 1 year ofage, from one month of age to less than four months of age, from threemonths of age to less than seven months of age, from six months of ageto less than 1 year of age, from 1 year of age to less than 2 years ofage, from 2 years of age to less than 3 years of age, from 2 years ofage to less than seven years of age, from 3 years of age to less than 5years of age, from 5 years of age to less than 10 years of age, from 6years of age to less than 13 years of age, from 10 years of age to lessthan 15 years of age, or from 15 years of age to less than 22 years ofage.

In one embodiment, provided herein is a method of treating a tumor, themethod comprising administering to a subject in need thereof atherapeutically effective amount of a compound of Formula I or apharmaceutically acceptable salt thereof or a compound of Formula II ora pharmaceutically acceptable salt thereof. In one embodiment, the tumoris a MEK-associated tumor. In one embodiment, the MEK-associated tumorhas a BRAF mutation. In one embodiment, the BRAF mutation is V600Eand/or V600K and/or V600D and/or V600R and/or V600S. In one embodiment,the BRAF mutation is V600E. In one embodiment, the BRAF mutation isV600K. In one embodiment, the MEK-associated tumor has a BRAF fusion,e.g., a BRAF fusion disclosed herein. In one embodiment, theMEK-associated tumor is a BRAF wild-type tumor.

In one embodiment of any of the methods of use described herein, thetumor is a solid tumor. In one embodiment of any of the methodsdisclosed herein, the solid tumor is a MEK-associated tumor. In oneembodiment, the tumor is intracranial. In one embodiment, the tumor isextracranial. In one embodiment of any of the methods of uses describedherein, the tumor (e.g., a MEK-associated tumor) is a malignant tumor(i.e., cancer) e.g., a MEK-associated cancer. In one embodiment of anyof the methods of use described herein, the MEK-associated cancer ismelanoma, colon cancer, colorectal cancer, lung cancer (e.g., small celllung carcinoma or non-small cell lung carcinoma), thyroid cancer (e.g.,papillary thyroid cancer, medullary thyroid cancer, differentiatedthyroid cancer, recurrent thyroid cancer, or refractory differentiatedthyroid cancer), breast cancer, ovarian cancer, cancer of the CNS, bonecancer, cancer of the anus, anal canal, or anorectum, cancer of the eye,bile duct cancer, ductal carcinoma in situ, liver cancer, gallbladder,or pleura, oral cancer, oral cavity cancer, lip cancer, oropharyngealcancer, cancer of the nose, nasal cavity or middle ear, cancer of thevulva, esophageal cancer, cervical cancer, gastrointestinal carcinoidtumor, hypopharynx cancer, kidney cancer, larynx cancer, liver cancer,lung cancer, melanoma, nasopharynx cancer, peripheral nervous systemcancers (e.g., neuroblastoma), ovarian cancer, pancreatic cancer,peritoneum, omentum, and mesentery cancer, pharynx cancer, prostatecancer, renal cancer (e.g., renal cell carcinoma (RCC)), small intestinecancer, soft tissue cancer, stomach cancer, testicular cancer, uterinecancer, ureter cancer, or urinary bladder cancer.

In one embodiment of any of the methods of use described herein, theMEK-associated cancer is an extracranial cancer (i.e., an extracranialtumor). In one embodiment, the extracranial cancer is selected frommelanoma, colorectal cancer, thyroid cancer, non-small cell lung cancer,ovarian cancer, and neuroblastoma. In one embodiment, the MEK-associatedcancer is melanoma. In one embodiment, the MEK-associated cancer iscolorectal cancer. In one embodiment, the MEK-associated cancer isthyroid cancer. In one embodiment, the MEK-associated cancer isnon-small cell lung cancer. In one embodiment, the MEK-associated canceris ovarian cancer. In one embodiment, the MEK-associated cancer isneuroblastoma.

In one embodiment of any of the methods of use described herein, theMEK-associated cancer is a CNS cancer.

In one embodiment of any of the methods of use described herein, theMEK-associated cancer is an intracranial cancer (brain cancer).

In one embodiment of any of the methods of use described herein, thecancer is a metastatic cancer.

The term “metastasis” is an art known term that refers to the spread ofcancer cells from the place where they first formed (the primary site)to one or more other sites in a subject (one or more secondary sites).In metastasis, cancer cells break away from the original (primary)tumor, travel through the blood or lymph system, and form a new tumor (ametastatic tumor) in other organs or tissues of the body. The new,metastatic tumor includes the same or similar cancer cells as theprimary tumor. At the secondary site, the tumor cell may proliferate andbegin the growth or colonization of a secondary tumor at this distantsite.

The term “metastatic cancer” (also known as “secondary cancer”) as usedherein refers to a type of cancer that originates in one tissue type,but then spreads to one or more tissues outside of the (primary)cancer's origin. Metastatic brain cancer refers to cancer in the brain,i.e., cancer which originated in a tissue other than the brain and hasmetastasized to the brain.

In one embodiment, compounds of Formula I or a pharmaceuticallyacceptable salts thereof and compounds of Formula II or apharmaceutically acceptable salts thereof exhibit surprising brainand/or CNS penetrance. Such compounds are capable of crossing the BBBand inhibiting a MEK kinase in the brain and/or other CNS structures.Accordingly, in one embodiment, compounds provided herein are useful fortreating a CNS tumor such as a CNS cancer.

In one embodiment, the MEK-associated tumor is a malignant CNS tumor(i.e., a MEK-associated CNS cancer). In one embodiment, theMEK-associated CNS cancer has a BRAF mutation. In one embodiment theMEK-associated CNS cancer is has a BRAF V600 mutation. In oneembodiment, the BRAF mutation is V600E and/or V600K and/or V600D and/orV600R and/or V600S. In one embodiment the MEK-associated CNS cancer ishas a BRAF V600E mutation. In one embodiment the MEK-associated CNScancer is has a BRAF V600K mutation. In one embodiment, theMEK-associated tumor has a BRAF fusion. In one embodiment, theMEK-associated tumor is a BRAF wild-type tumor.

The term “CNS cancer” or “cancer of the CNS” as used interchangeablyherein refers to a cancer (i.e., a malignant tumor) of the CNS,including cancers of the brain (also known as intracranial tumors),cancers of the spinal cord, and cancers of the meninges surrounding thebrain and spinal cord. Cancers of the brain include metastatic braincancers (i.e., metastatic intracranial cancers) and malignant primarybrain tumors.

In one embodiment, the MEK-associated CNS cancer is a MEK-associatedmetastatic brain cancer. The MEK-associated metastatic brain cancer maybe the result of any cancer described herein, wherein the subject hasdeveloped at least one brain metastasis. In one embodiment, themetastatic brain cancer is melanoma, colorectal cancer, thyroid cancer,non-small cell lung cancer, ovarian cancer, or neuroblastoma. In oneembodiment, the MEK-associated metastatic brain cancer is metastaticmelanoma, metastatic colorectal cancer, or metastatic non-small celllung cancer. In one embodiment, the MEK-associated metastatic braincancer is metastatic melanoma. In one embodiment, the MEK-associatedmetastatic brain cancer is metastatic colorectal cancer. In oneembodiment, the MEK-associated metastatic brain cancer is metastaticnon-small cell lung cancer. In one embodiment, the MEK-associatedmetastatic brain cancer is metastatic ovarian cancer. In one embodiment,the metastatic brain cancer is metastatic thyroid cancer. In oneembodiment, the MEK-associated metastatic brain cancer is kidney cancer.In one embodiment, the cancer is MEK-associated metastatic cancer withat least one brain metastasis (i.e., a metastatic brain cancer). In oneembodiment, the cancer is MEK-associated metastatic melanoma with atleast one brain metastasis. In one embodiment, the cancer isMEK-associated metastatic colorectal cancer with at least one brainmetastasis. In one embodiment, the cancer is MEK-associated metastaticnon-small cell lung cancer with at least one brain metastasis. In oneembodiment, the cancer is MEK-associated metastatic ovarian cancer withat least one brain metastasis. In one embodiment, the cancer isMEK-associated metastatic thyroid cancer with at least one brainmetastasis. In one embodiment, the cancer is MEK-associatedneuroblastoma with at least one brain metastasis. In one embodiment ofany of said MEK-associated metastatic brain cancers, the cancer has aBRAF mutation. In one embodiment the cancer has a BRAF V600 mutation. Inone embodiment, the BRAF mutation is V600E and/or V600K and/or V600Dand/or V600R and/or V600S. In one embodiment the cancer is has a BRAFV600E mutation. In one embodiment the cancer is has a BRAF V600Kmutation. In one embodiment, the MEK-associated tumor has a BRAF fusion.In one embodiment, the MEK-associated tumor is a BRAF wild-type tumor.

In one embodiment, the MEK-associated cancer is leptomeningealmetastases (leptomeningeal disease (LMD)). LMD represents a subset ofCNS metastases that grow in the lining of the brain or spine and/or inthe cerebrospinal fluid (CSF), or leptomeningeal carcinomatosis. Inmammals, the meninges are the dura mater, the arachnoid mater, and thepia mater. CSF is located in the subarachnoid space between thearachnoid mater and the pia mater. The arachnoid and pia mater togetherare sometimes called the leptomeninges. When LMD occurs in theleptomeninges and/or CSF surrounding the spinal cord, it may be referredto as “extracranial LMD”. When LMD occurs in the leptomeninges and/orCSF of the brain, it may be referred to as “intracranial LMD”. Since LMDcancer cells can be suspended in the CSF, they can quickly spreadthroughout the CNS. As a result, LMD has a poor prognosis, with survivaltypically measured in months. In one embodiment, the metastatic canceris LMD. In one embodiment, the metastatic cancer is a MEK-associatedLMD. In one embodiment, the metastatic cancer is a MEK-associatedintracranial LMD. In one embodiment, the metastatic cancer is aMEK-associated extracranial LMD. In one embodiment the MEK-associatedLMD is LMD derived from melanoma metastases (i.e., the LMD is metastaticmelanoma). In one embodiment the MEK-associated LMD is LMD derived fromcolorectal cancer metastases (i.e., the LMD is metastatic colorectalcancer). In one embodiment the MEK-associated LMD is LMD derived fromnon-small cell lung cancer metastases (i.e., the LMD is metastaticnon-small cell lung cancer). In one embodiment of any of said theMEK-associated LMD's, the LMD has a BRAF mutation. In one embodiment theMEK-associated LMD has a BRAF V600 mutation. In one embodiment, the BRAFmutation is V600E and/or V600K and/or V600D and/or V600R and/or V600S.In one embodiment the MEK-associated LMD is has a BRAF V600E mutation.In one embodiment the MEK-associated LMD is has a BRAF V600K mutation.In one embodiment, the MEK-associated LMD has a BRAF fusion. In oneembodiment, the MEK-associated LMD is a BRAF wild-type tumor.

In one embodiment, the MEK-associated tumor is a cancer having a highrisk of metastasis. In one embodiment, the tumor having a high risk ofmetastasis is a cancer having a BRAF V600E, V600D, V600K, V600R and/orV600S mutation. In one embodiment, the cancer having a high risk ofmetastasis has a BRAF fusion, e.g., any of the BRAF fusions disclosedherein. In one embodiment, the cancer having a high risk of metastasisis a BRAF wild-type tumor. In one embodiment, the cancer having a highrisk of metastasis is melanoma, colorectal cancer, thyroid cancer,non-small cell lung cancer, ovarian cancer or neuroblastoma. In oneembodiment, the cancer having a high risk of metastasis is melanoma,colorectal cancer, thyroid cancer, non-small cell lung cancer, ovariancancer or neuroblastoma. In one embodiment, the cancer having a highrisk of metastasis is melanoma. In one embodiment, the cancer having ahigh risk of metastasis is melanoma having a BRAF V600E mutation or BRAFV600K mutation. In one embodiment, the cancer having a high risk ofmetastasis is colorectal cancer. In one embodiment, the cancer having ahigh risk of metastasis is colorectal cancer having a BRAF V600Emutation or BRAF V600K mutation. In one embodiment, the cancer having ahigh risk of metastasis is thyroid cancer. In one embodiment, the cancerhaving a high risk of metastasis is thyroid cancer having a BRAF V600Emutation or BRAF V600K mutation. In one embodiment, the cancer having ahigh risk of metastasis is non-small cell lung cancer. In oneembodiment, the cancer having a high risk of metastasis is non-smallcell lung cancer having a BRAF V600E mutation or BRAF V600K mutation. Inone embodiment, the cancer having a high risk of metastasis is ovariancancer. In one embodiment, the cancer having a high risk of metastasisis ovarian cancer having a BRAF V600E mutation or BRAF V600K mutation.In one embodiment, the cancer having a high risk of metastasis isneuroblastoma. In one embodiment, the cancer having a high risk ofmetastasis is neuroblastoma having a BRAF V600E mutation or BRAF V600Kmutation. In one embodiment, the cancer having a high risk of metastasishas a KIAA11549-BRAF fusion.

In one embodiment, the CNS tumor is a primary brain tumor. Primary braintumors are tumors that start in the brain or spine and are knowncollectively as gliomas. The term “glioma” is used to describe tumorsthat originate in glial cells present in the CNS. According to the WHOclassification of brain tumors, gliomas are graded by the cell activityand aggressiveness on a scale including Grade I (benign CNS tumors) andGrades I to IV (malignant CNS tumors):

Grade I glioma (Pilocytic astrocytoma): typically occurs in children inthe cerebellum or brainstem, and occasionally in the cerebralhemispheres, and are slow growing. Grade I can occur in adults. Althoughthey are benign (WHO grade 1), the difficulty in curing this diseasemakes their growth malignant in behavior with high morbidity rates(Rostami, Acta Neurochir (Wien). 2017; 159(11): 2217-2221).

Grade II glioma (Low-grade gliomas): includes astrocytoma,oligodendroglioma, and mixed oligoastrocytoma. Grade II gliomastypically occur in young adults (e.g., ages 20-50) and are most oftenfound in the cerebral hemispheres. Due to the infiltrative nature ofthese tumors, recurrences may occur. Some grade II gliomas recur andevolve into more aggressive tumors (grade III or IV).

Grade III glioma (Malignant glioma): includes anaplastic astrocytoma,anaplastic oligodendroglioma, and anaplastic mixed oligoastrocytoma.Grade III tumors are aggressive, high-grade cancers and invade nearbybrain tissue with tentacle-like projections, making complete surgicalremoval more difficult.

Grade IV gliomas: includes Glioblastoma multiforme (GBM) andgliosarcoma; (GBM) is a malignant glioma. GBM is the most aggressive andmost common primary brain tumor. Glioblastoma multiforme usually spreadsquickly and invades other parts of the brain, with tentacle-likeprojections, making complete surgical removal more difficult.Gliosarcoma is a malignant cancer and is defined as a glioblastomaconsisting of gliomatous and sarcomatous components.

In one embodiment, the primary brain tumor is a glioma. In oneembodiment, the glioma is a low-grade glioma. In one embodiment, theglioma is a pediatric low-grade glioma.

In one embodiment, the primary brain tumor is a benign primary braintumor. Benign primary brain tumors can cause severe pain, permanentbrain damage and death, and in some cases, become malignant.Non-limiting examples of benign primary brain tumors include Grade Igliomas, papillary craniopharyngiomas, meningioma (including rhabdoidmeningioma), atypical teratoid/rhabdoid tumors, and dysembryoplasticneuroepithelial tumor (DNT), pilocytic astrocytoma, oligodendroglioma,mixed oligoastrocytoma, anaplastic astrocytoma, anaplasticoligodendroglioma, anaplastic mixed oligoastrocytoma, diffuseastrocytoma, ependymoma, a pleomorphic xanthoastrocytoma (PXA), aganglioglioma, a gliosarcoma, or an anaplastic ganglioglioma.

In one embodiment, the cancer is a peripheral nervous system cancer. Inone embodiment, the peripheral nervous system cancer is neuroblastoma.

In one embodiment, provided herein is a method of treating aMEK-associated CNS tumor, comprising administering (e.g., oraladministration) to a subject in need thereof a therapeutically effectiveamount of a compound of Formula I or a pharmaceutically acceptable saltthereof or a compound of Formula II or a pharmaceutically acceptablesalt thereof to the subject in need thereof. In one embodiment, theMEK-associated CNS tumor has a BRAF V600 mutation. In one embodiment,the MEK-associated CNS tumor has a BRAF V600E and/or V600K and/or V600Dand/or V600R mutation and/or V600S. In one embodiment, theMEK-associated CNS tumor has a BRAF V600E mutation. In one embodiment,the MEK-associated CNS tumor has a BRAF V600K mutation. In oneembodiment, the MEK-associated CNS tumor has a BRAF fusion, e.g., any ofthe BRAF fusions disclosed herein, e.g., a KIAA11549-BRAF fusion. In oneembodiment, the MEK-associated CNS tumor is a BRAF wild-type tumor. Inone embodiment, the subject has been treated with one or more,anticancer therapies independently selected from anticancer agents,surgery and radiotherapy prior to administration of a compound ofFormula I or a pharmaceutically acceptable salt thereof or a compound ofFormula II or a pharmaceutically acceptable salt thereof, e.g., asdescribed hereinbelow. In one embodiment, the subject is treated with atherapeutically effective amount compound of a compound of Formula I ora pharmaceutically acceptable salt thereof or a compound of Formula IIor a pharmaceutically acceptable salt thereof in combination with one ormore anticancer therapies independently selected from one or moreanticancer agents, surgery and/or radiotherapy, e.g., as describedhereinbelow. In one embodiment, the subject is treated with one or moreanticancer therapies, independently selected from an anticancer agent,surgery, and radiotherapy after administration of a compound of FormulaI or a pharmaceutically acceptable salt thereof or a compound of FormulaII or a pharmaceutically acceptable salt thereof, e.g., as describedhereinbelow. In one embodiment, the MEK-associated tumor is a CNS tumor.In one embodiment, the MEK-associated CNS tumor is a malignant CNS tumor(i.e., a CNS cancer). In one embodiment, the malignant CNS tumor is ametastatic CNS cancer. In one embodiment, the metastatic CNS cancer isselected from metastatic melanoma, metastatic colorectal cancer,metastatic non-small cell lung cancer, metastatic thyroid cancer, andmetastatic ovarian cancer. In one embodiment, the metastatic CNS canceris metastatic melanoma. In one embodiment, the metastatic CNS cancer iscolorectal cancer. In one embodiment, the metastatic CNS cancer ismetastatic non-small cell lung cancer. In one embodiment, the metastaticCNS cancer is metastatic thyroid cancer. In one embodiment, themetastatic CNS cancer is metastatic ovarian cancer. In one embodiment,the MEK-associated CNS cancer is LMD. In one embodiment, the LMD isintracranial. In one embodiment, the LMD is extracranial. In oneembodiment, the LMD is metastatic melanoma. In one embodiment, the LMDis selected from metastatic melanoma, metastatic colorectal cancer, andmetastatic non-small cell lung cancer. In one embodiment, the LMD ismetastatic colorectal cancer. In one embodiment, the LMD is metastaticnon-small cell lung cancer. In one embodiment, the MEK-associated CNScancer is a primary brain tumor. In one embodiment, the primary braintumor is a Grade 2 glioma. In one embodiment, the primary brain tumor isa Grade 3 glioma. In one embodiment, the primary brain tumor is a Grade4 glioma. In one embodiment, the MEK-associated CNS tumor is a benigntumor. In one embodiment, the benign CNS tumor is a papillarycraniopharyngioma, a meningioma (including rhabdoid meningioma), anatypical teratoid/rhabdoid tumor, or a dysembryoplastic neuroepithelialtumor (DNT). In one embodiment, the compound is a compound of Formula Ior a pharmaceutically acceptable salt thereof. In one embodiment, thecompound is a compound of Formula II or a pharmaceutically acceptablesalt thereof. In one embodiment, the compound of is selected from acompound of Examples 1-69 or a pharmaceutically acceptable salt thereof.

The ability to determine whether a compound may be suitable for treatinga CNS cancer may be determined, for example, by identifying if thecompound is a substrate of an efflux transporter and/or measuring thecell permeability and/or measuring the free blood-to-free plasma ratio,as described herein.

In one embodiment, compounds of Formula I or pharmaceutically acceptablesalts thereof and compounds of Formula II or pharmaceutically acceptablesalt thereof, exhibit high cell permeability. Methods for determiningthe permeability of a compound of the invention can be determinedaccording to the assay described in Example B, and permeabilitycoefficients are provided in Table B1.

Compounds of the invention exhibit low efflux. In vitro methods ofevaluating whether compounds of the invention are substrates for theefflux transporters P-glycoprotein (P-gp or Multi-drug Resistance 1(MDR1) protein) and Breast cancer resistance protein (BCRP) aredescribed in Example B, and efflux ratios of compounds of the inventionare provided in Table B3.

In one embodiment, compounds of the invention exhibit medium-to-highbrain (unbound)/plasma (unbound) ratios (i.e., medium-to-high freebrain/plasma ratios). The ability of a compound of the invention topenetrate the BBB of a subject (e.g., a human) can be determined in asuitable animal model (e.g., a rodent, such as a mouse). For example,the ability of certain compounds to penetrate the BBB in mice wasdetermined by evaluating the unbound brain-to-unbound plasmaconcentration (free B/P) ratio in mice e.g. as described in Example C,and the free brain-to-free plasma ratios are provided in Table C2.Calculating the free brain-to-free plasma ratio of compounds enablespredictions of efficacious concentrations required to achieve efficacyin the periphery and the brain based upon dose-related exposure inanimal models. These distribution data, along with the associatedpharmacokinetic data, can be utilized to model and predict the doserequired to achieve efficacy in human patients.

Accordingly, in one embodiment, the methods of the present inventioninclude methods for treating a MEK-associated CNS cancer in a subject inneed thereof, comprising administering a compound of Formula II orpharmaceutically acceptable salt, wherein at least a portion of thecompound of Formula II penetrates the BBB, as demonstrated in a suitableanimal model. In one embodiment, the brain/plasma ratio of total drug isat least about 0.3 after administration (e.g., oral or intravenousadministration) to a subject. In one embodiment, the brain/plasma ratioof total drug is at least about 0.35 after administration (e.g., oral orintravenous administration) to a subject. In one embodiment, thebrain/plasma ratio of total drug is at least about 0.4 afteradministration (e.g., oral or intravenous administration) to a subject.In one embodiment, the brain/plasma ratio of total drug is at leastabout 0.45 after administration (e.g., oral or intravenousadministration) to a subject. In one embodiment, the brain/plasma ratioof total drug is at least about 0.5 after administration (e.g., oral orintravenous administration) to a subject. In one embodiment, thebrain/plasma ratio of total drug is at least about 0.55 afteradministration (e.g. oral or intravenous administration) to a subject.In one embodiment, the brain/plasma ratio of total drug is at leastabout 0.6 after administration (e.g. oral or intravenous administration)to a subject. In one embodiment, the brain/plasma ratio of total drug isat least about 0.65 after administration (e.g. oral or intravenousadministration) to a subject. In one embodiment, the brain/plasma ratioof total drug is at least about 0.7 after administration (e.g. oral orintravenous administration) to a subject. In one embodiment, thebrain/plasma ratio of total drug is at least about 0.75 afteradministration (e.g. oral or intravenous administration) to a subject.In one embodiment, the brain/plasma ratio of total drug is at leastabout 0.8 after administration (e.g. oral or intravenous administration)to a subject. In one embodiment, the brain/plasma ratio of total drug isat least about 0.85 after administration (e.g. oral or intravenousadministration) to a subject. In one embodiment, the brain/plasma ratioof total drug is at least about 0.9 after administration (e.g. oral orintravenous administration) to a subject. In one embodiment, thebrain/plasma ratio of total drug is at least about 0.95 afteradministration (e.g. oral or intravenous administration) to a subject.In one embodiment, the brain/plasma ratio of total drug is at leastabout 1.0 after administration (e.g. oral or intravenous administration)to a subject. In one embodiment, the brain/plasma ratio of total drug isat least about 1.0 after administration (e.g. oral or intravenousadministration) to a subject. In one embodiment, the brain/plasma ratioof total drug is at least about 1.1 after administration (e.g. oral orintravenous administration) to a subject. In one embodiment, thebrain/plasma ratio of total drug is at least about 1.2 afteradministration (e.g. oral or intravenous administration) to a subject.In one embodiment, the brain/plasma ratio of total drug is at leastabout 1.3 after administration (e.g. oral or intravenous administration)to a subject. In one embodiment, the brain/plasma ratio of total drug isat least about 1.4 after administration (e.g. oral or intravenousadministration) to a subject. In one embodiment, the brain/plasma ratioof total drug is at least about 1.5 after administration (e.g. oral orintravenous administration) to a subject. In one embodiment, thebrain/plasma ratio of total drug is at least about 1.6 afteradministration (e.g. oral or intravenous administration) to a subject.In one embodiment, the brain/plasma ratio of total drug is at leastabout 1.7 after administration (e.g. oral or intravenous administration)to a subject. It is to be noted that the percentage of a compound thatpenetrates the BBB is calculated based upon the area under theconcentration-time curve for a given time period (AUC_(0-t)) in thebrain versus the plasma. Accordingly, the percentages represent a ratioof concentrations. That is, if (AUC₀₋₂₄ h) for a compound is 20 ng/mL inthe brain and 80 ng/mL in the plasma, then the percentage of thecompound that penetrates the BBB is 20% (20 ng/mL in the brain dividedby the total concentration of (20 ng/mL+80 ng/mL)) (i.e., abrain-to-plasma ratio of 0.20). In one embodiment, the percentages arecalculated based upon the area under the concentration-time curve forthe time period from t=0 (time of dosing) to the last quantifiableconcentration point, i.e., (AUC_(0-last)).

Cancers that frequently metastasize to the brain are known to carry MAPKpathway activating alterations such as BRAF mutations, including theBRAF mutations disclosed herein, or BRAF fusions, including the BRAFfusions disclosed herein. Although activating mutations can occur atdifferent levels in the canonical pathway, they all require signalingvia mitogen/extracellular signal-regulated kinase (MEK) in order toincrease proliferation and survival (Schubbert S, Shannon K, Bollag G.Nat Rev Cancer. 2007; 7:295-308). Mutations in the BRAF gene have beenidentified in malignant melanomas, papillary thyroid carcinomas,colorectal carcinomas, non-small cell lung carcinoma (NSCLC), andovarian carcinomas and metastatic tumors thereof, and in primary braintumors (Davies H., et al., Nature 417(6892):949-954, 2002). For example,BRAF mutations, such as BRAF V600 mutations, have been observed innumerous metastatic CNS tumors, including melanoma brain metastases(Flaherty K T, et al., Nat Rev Cancer (2012) 12(5):349-61), brainmetastases of colorectal cancers and brain metastases of non-small celllung cancer (Berghoff, A S, Preusser M., Curr Opin Neurol (2014)27(6):689-696), papillary thyroid cancer (Kim, W W et al., J OtolaryngolHead Neck Surg. 2018; 47:4, 1-6), and ovarian cancer (Grisham R N., etal., Cancer, 2013; 119:548-554).

BRAF mutations, e.g., the BRAF mutations disclosed herein, and BRAFfusions, e.g., the BRAF fusions disclosed herein, have also beenobserved in malignant primary brain tumors, including Grade IV gliomas,e.g., glioblastomas and gliosarcomas, anaplastic astrocytomas(high-grade tumors) and WHO grade III anaplastic gangliogliomas(Berghoff, A S, Preusser M., Curr Opin Neurol (2014) 27(6):689-696);Schindler et al. (Acta Neuropathol 121(3):397-405, 2011); Behling et al.(Diagn Pathol 11(1):55, 2016); K. C. Schreck, et al., Cancers, 2019, 11,1262)) in pediatric and adult populations.

BRAF mutations, e.g., the BRAF mutations disclosed herein, and BRAFfusions, e.g., the BRAF fusions disclosed herein, have also beenobserved in benign primary brain tumors, for example in WHO Grade IIastrocytomas, WHO grade II pleomorphic xanthoastrocytomas (PXAs),pleomorphic xanthoastrocytomas with anaplasia, Pilocytic astrocytoma(PA), papillary craniopharyngiomas, gangliogliomas, astroblastomas,pilocytic astrocytomas, atypical teratoid/rhabdoid tumors, rhabdoidmeningiomas (Berghoff, A S, Preusser M., Curr Opin Neurol (2014)27(6):689-696; Schindler et al. (Acta Neuropathol 121(3):397-405, 2011);Behling et al. (Diagn Pathol 11(1):55, 2016); (Behling et al., DiagnPathol 11(1):55, 2016; Brastianos et al., Nat Genet 46(2):161-165, 2014;Dougherty et al., Neuro Oncol 12(7):621-630, 2010; Lehman et al., NeuroOncol 19(1):31-42, 2017; Mordechai et al., Pediatr Hematol Oncol32(3):207-211, 2015; Myung et al., Transl Oncol 5(6):430-436, 2012;Schindler et al., Acta Neuropathol 121(3):397-405, 2011)), in pediatricand adult populations.

BRAF mutations have also been detected in relapsed neuroblastomas(Eleveld, T F, et al., Nat Genet 47(8):864-871, 2015). Neuroblastoma isa pediatric tumor of the peripheral nervous system. The majority ofneuroblastoma subjects have tumors that initially respond tochemotherapy, but a large proportion of subjects will experiencetherapy-resistant relapses.

Accordingly, also provided herein is a method for treating a subjectdiagnosed with or identified as having a MEK-associated tumor, e.g., anyof the exemplary MEK-associated tumors disclosed herein, comprisingadministering to the subject a therapeutically effective amount of acompound of Formula I or a pharmaceutically acceptable salt thereof or acompound of Formula II or a pharmaceutically acceptable salt thereof,wherein the subject that has been identified or diagnosed as having atumor having a BRAF mutation of BRAF fusion, e.g., through the use of aregulatory agency-approved, e.g., FDA-approved test or assay foridentifying BRAF mutation or fusion in a subject or a biopsy sample fromthe subject or by performing any of the non-limiting examples of assaysdescribed herein. In one embodiment, the test or assay is provided as akit. In one embodiment, the assay utilizes next generation sequencing,pyrosequencing, immunohistochemistry, fluorescence microscopy, breakapart FISH analysis, Southern blotting, Western blotting, FACS analysis,Northern blotting, or PCR-based amplification (e.g., RT-PCR andquantitative real-time RT-PCR). In one embodiment, the assay is aregulatory agency-approved assay, e.g., FDA-approved kit.

In one embodiment, the biopsy is a tumor biopsy (e.g., a tumor sampleobtained during traditional surgery or a stereotactic needle biopsy,e.g., a stereotactic need biopsy guided by CT or MRI scanning). Tissuebiopsy methods can be used to detect total tumor burden and/or the BRAFmutation and/or the BRAF fusion.

In one embodiment, the BRAF mutation or fusion can be identified using aliquid biopsy (variously referred to as a fluid biopsy or fluid phasebiopsy). See, e.g., Karachialiou et al., “Real-time liquid biopsiesbecome a reality in cancer treatment”, Ann. Transl. Med., 3(3):36, 2016.Liquid biopsy methods can be used to detect total tumor burden and/orthe BRAF mutation. Liquid biopsies can be performed on biologicalsamples obtained relatively easily from a subject (e.g., via a simpleblood draw) and are generally less invasive than traditional methodsused to detect tumor burden and/or BRAF mutation. In one embodiment,liquid biopsies can be used to detect the presence of a BRAF mutation atan earlier stage than traditional methods. In one embodiment, thebiological sample to be used in a liquid biopsy can include CSF, blood,plasma, urine, saliva, sputum, broncho-alveolar lavage, bile, lymphaticfluid, cyst fluid, stool, ascites, and combinations thereof. In oneembodiment, a liquid biopsy can be used to detect circulating tumorcells (CTCs). In one embodiment, a liquid biopsy can be used to detectcell-free DNA. In one embodiment, the cell-free DNA detected using aliquid biopsy is circulating tumor DNA (ctDNA) that is derived fromtumor cells. Analysis of ctDNA (e.g., using sensitive detectiontechniques such as, without limitation, next-generation sequencing(NGS), traditional PCR, digital PCR, or microarray analysis) can be usedto identify a BRAF mutation or BRAF fusion.

In one embodiment, a BRAF mutation or BRAF fusion identified using aliquid biopsy is also present in a cancer cell that is present in thesubject (e.g., in a tumor). In one embodiment, any of the types of BRAFmutations or fusions can be detected using a liquid biopsy. In oneembodiment, a genetic mutation identified via a liquid biopsy can beused to identify the subject as a candidate for a particular treatment.

“Tumor burden” also referred to as “tumor load”, refers to the totalamount of tumor material distributed throughout the body. Tumor burdenrefers to the total number of cancer cells or the total size oftumor(s), throughout the body, including lymph nodes and bone narrow.Tumor burden can be determined by a variety of methods known in the art,such as, e.g. by measuring the dimensions of tumor(s) upon removal fromthe subject, e.g., using calipers, or while in the body using imagingtechniques, e.g., magnetic resonance imaging (MRI) scans, computedtomography (CT), multi-detector CT (MDCT), positron emission tomography(PET), X-ray, ultrasound, or bone scan.

The term “tumor size” refers to the total size of the tumor which can bemeasured as the length and width of a tumor. Tumor size may bedetermined by a variety of methods known in the art, such as, e.g., bymeasuring the dimensions of tumor(s) upon removal from the subject,e.g., using calipers, or while in the body using imaging techniques,e.g., MRI scans, bone scan, ultrasound, or CT.

Liquid biopsies can be performed at multiple times during a course ofdiagnosis, a course of monitoring, and/or a course of treatment todetermine one or more clinically relevant parameters including, withoutlimitation, progression of the disease or efficacy of a treatment, afteradministering a treatment to the subject. For example, a first liquidbiopsy can be performed at a first time point and a second liquid biopsycan be performed at a second time point during a course of diagnosis, acourse of monitoring, and/or a course of treatment. In one embodiment,the first time point can be a time point prior to diagnosing a subjectwith a disease (e.g., when the subject is healthy), and the second timepoint can be a time point after subject has developed the disease (e.g.,the second time point can be used to diagnose the subject with thedisease). In one embodiment, the first time point can be a time pointprior to diagnosing a subject with a disease (e.g., when the subject ishealthy), after which the subject is monitored, and the second timepoint can be a time point after monitoring the subject. In oneembodiment, the first time point can be a time point after diagnosing asubject with a disease, after which a treatment is administered to thesubject, and the second time point can be a time point after thetreatment is administered; in such cases, the second time point can beused to assess the efficacy of the treatment (e.g., if the geneticmutation(s) detected at the first time point are reduced in abundance orare undetectable).

A compound of Formula I or a pharmaceutically acceptable salt thereof ora compound of Formula II or a pharmaceutically acceptable salt thereofmay be used alone or in combination with one or more different forms oftreatment to treat a subject with an abnormal cell growth, such as aMEK-associated tumor, for example a MEK-associated cancer.

In one embodiment, a compound of Formula I or a pharmaceuticallyacceptable salt thereof or a compound of Formula II or apharmaceutically acceptable salt thereof may be used in combination withone or more, e.g., one or more additional anticancer therapies, forexample one or more therapies independently selected from surgery,radiotherapy and anticancer agents that work by the same or by adifferent mechanism of action. In one embodiment, treatment of a subjecthaving a MEK-associated cancer with a compound of Formula I or apharmaceutically acceptable salt thereof or a compound of Formula II ora pharmaceutically acceptable salt thereof in combination with one ormore, e.g., one or more additional therapies, e.g., independentlyselected from one or more of surgery, radiotherapy, and anticanceragents (e.g., any of the anticancer agents described herein below,wherein the anticancer agent is other than a compound of Formula I or apharmaceutically acceptable salt thereof or a compound of Formula II ora pharmaceutically acceptable salt thereof), can have increasedtherapeutic efficacy as compared to treatment of the same subject or asimilar subject with a compound of Formula I or a pharmaceuticallyacceptable salt thereof or a compound of Formula II or apharmaceutically acceptable salt thereof as a monotherapy. When acombination therapy is used and the one or more, e.g., one two or threeanticancer therapies are independently selected from one or moreanticancer agents such as any of the anticancer agents disclosed herein,the anticancer agent(s) may be administered simultaneously or separatelywith variable intervening time limits and in any order with a compoundof the invention in any order and using varying dosing schedules. In oneembodiment, the anticancer agent(s) is administered to the subject priorto administration of the compound of the invention. In anotherembodiment, the anticancer agent(s) is administered to the subject afteradministration of the compound of the invention. In another embodiment,the anticancer agent(s) is administered to the subject simultaneouslywith the administration of the compound of the invention. In oneembodiment, a compound of Formula I or a pharmaceutically acceptablesalt thereof or a compound of Formula II or a pharmaceuticallyacceptable salt thereof is used in combination with one additionalanticancer therapy which is surgery, radiotherapy, or an anticanceragent that work by the same or by a different mechanism of action.

Accordingly, in one embodiment, provided herein are methods of treatinga subject having a MEK-associated tumor (e.g., any of the MEK-associatedtumors described herein) that comprise administering to the subject atherapeutically effective amount of a compound of Formula I or apharmaceutically acceptable salt thereof or a compound of Formula II ora pharmaceutically acceptable salt thereof in combination with one ormore additional anticancer therapies. In one embodiment, the anticancertherapy is one or more anticancer agents other than a compound ofFormula I or a pharmaceutically acceptable salt thereof or a compound ofFormula II or a pharmaceutically acceptable salt thereof. In oneembodiment, the anticancer therapy is one anticancer agent other than acompound of Formula I or a pharmaceutically acceptable salt thereof or acompound of Formula II or a pharmaceutically acceptable salt thereof. Inone embodiment, the additional anticancer therapy is surgery. In oneembodiment, the additional anticancer therapy is radiotherapy.

Also provided herein is a compound of Formula I or a pharmaceuticallyacceptable salt thereof or a compound of Formula II or apharmaceutically acceptable salt thereof for use in combination with oneor more, e.g., one or more anticancer therapies. In one embodiment, theadditional anticancer therapy is independently selected from one or moretherapies independently selected from surgery, radiotherapy, and/or oneor more anticancer agents that work by the same or by a differentmechanism of action.

Also provided herein is one or more, e.g., one or more anticancertherapies for use in combination with a compound of Formula I or apharmaceutically acceptable salt thereof or a compound of Formula II ora pharmaceutically acceptable salt thereof. In one embodiment, theadditional anticancer therapy is independently selected from one or moretherapies independently selected from surgery, radiotherapy, and/or oneor more anticancer agents that work by the same or by a differentmechanism of action.

Also provided herein is a compound of Formula I or a pharmaceuticallyacceptable salt thereof or a compound of Formula II or apharmaceutically acceptable salt thereof for use in treating aMEK-associated tumor in combination with one or more, e.g., one or moreadditional anticancer therapies.

Also provided herein is one or more, e.g., one or more additionalanticancer therapies, for use in treating a MEK-associated tumor byco-administration with a compound of a compound of Formula I or apharmaceutically acceptable salt thereof or a compound of Formula II ora pharmaceutically acceptable salt thereof.

In one embodiment for treating a subject having a MEK-associated tumor,the subject is administered one or more anticancer therapies other thana compound of Formula I or a pharmaceutically acceptable salt thereof ora compound of Formula II or a pharmaceutically acceptable salt thereof,prior to administration of a compound of Formula I or a pharmaceuticallyacceptable salt thereof or a compound of Formula II or apharmaceutically acceptable salt thereof. In one embodiment, the one ormore anticancer therapies is selected from surgery, radiotherapy, and ananticancer agent that work by the same or by a different mechanism ofaction. For example, in one embodiment, a subject in need thereof mayundergo at least partial resection of the tumor prior to administrationof a compound of Formula I or a pharmaceutically acceptable salt thereofor a compound of Formula II or a pharmaceutically acceptable saltthereof. In one embodiment, the treatment by at least partial resectionof the tumor reduces the size of the tumor (e.g., the tumor burden)occurs prior to administration of one or more doses of a compound ofFormula I or a pharmaceutically acceptable salt thereof or a compound ofFormula II or a pharmaceutically acceptable salt thereof. In oneembodiment, a subject in need thereof may undergo radiotherapy prior toadministration of a compound of Formula I or a pharmaceuticallyacceptable salt thereof or a compound of Formula II or apharmaceutically acceptable salt thereof. In one embodiment, a subjectin need thereof may undergo treatment with one or more anticancer agentsother than a compound of Formula I or a pharmaceutically acceptable saltthereof or a compound of Formula II or a pharmaceutically acceptablesalt thereof prior to administration of a compound of Formula I or apharmaceutically acceptable salt thereof or a compound of Formula II ora pharmaceutically acceptable salt thereof. In one embodiment, a subjecthas a cancer that is refractory or intolerant to the previous therapy ortherapies.

Accordingly, in one embodiment, provided herein are methods of treatinga subject having a MEK-associated tumor, comprising (i) administeringone or more, e.g., one or more anticancer therapies to said subject, and(ii) after (i), administering (a) a compound of Formula I or apharmaceutically acceptable salt thereof or a compound of Formula II ora pharmaceutically acceptable salt thereof as monotherapy or (b) acompound of Formula I or a pharmaceutically acceptable salt thereof or acompound of Formula II or a pharmaceutically acceptable salt thereof incombination with one or more, e.g., one or more additional anticancertherapies. In one embodiment, the additional anticancer therapy isindependently selected from one or more of surgery, radiotherapy, and/orone or more anticancer agents that work by the same or by a differentmechanism of action. In one embodiment, the additional anticancertherapy is one or more anticancer agents that work by the same or by adifferent mechanism of action. In one embodiment, the additionalanticancer therapy is one anticancer agent that works by the same or bya different mechanism of action. In one embodiment, the additionalanticancer therapy is surgery. In one embodiment, the additionalanticancer therapy is radiotherapy.

Non-limiting examples of additional anticancer agents that can be usedin combination with a compound of Formula I or a pharmaceuticallyacceptable salt thereof or a compound of Formula II or apharmaceutically acceptable salt thereof according to any of thecombination therapy methods described herein include additional kinaseinhibitors other than a compound of Formula I or a pharmaceuticallyacceptable salt thereof or a compound of Formula II or apharmaceutically acceptable salt thereof, including MEK inhibitors, BRAFinhibitors, EGFR inhibitors, inhibitors of HER2 and/or HER3, SHP2inhibitors, Axl inhibitors, PI3K inhibitors, SOS1 inhibitors, signaltransduction pathway inhibitors, checkpoint inhibitors, modulators ofthe apoptosis pathway, cytotoxic chemotherapeutics,angiogenesis-targeted therapies, and immune-targeted agents includingimmunotherapy.

In one embodiment, the anticancer agent that can be used in combinationwith a compound of Formula I or a pharmaceutically acceptable saltthereof or a compound of Formula II or a pharmaceutically acceptablesalt thereof according to any of the combination therapy methodsdescribed herein is a targeted therapeutic agent. A “targetedtherapeutic agent” as used herein includes, refers to a molecule thatblocks the growth of cancer cells by interfering with specific targetedmolecules needed for carcinogenesis and tumor growth, rather than bysimply interfering with all rapidly dividing cells (e.g., withtraditional cytotoxic chemotherapy), and includes but is not limited to,receptor tyrosine kinase-targeted therapeutic agents, signaltransduction pathway inhibitors (for example, Ras-Raf-MEK-ERK pathwayinhibitors, PI3K-Akt-mTOR-S6K pathway inhibitors (“PI3K inhibitors”)),and modulators of the apoptosis pathway.

In one embodiment, the anticancer agent that can be used in combinationwith a compound of Formula I or a pharmaceutically acceptable saltthereof or a compound of Formula II or a pharmaceutically acceptablesalt thereof according to any of the combination therapy methodsdescribed herein is a BRAF inhibitor. Non-limiting examples of otherBRAF inhibitors include encorafenib, dabrafenib, vemurafenib,N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide(PLX4720), and(3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide(PLX8394), and pharmaceutically acceptable salts thereof, the compoundsdisclosed in International Application No. PCT/IB2020/055992, publishedDec. 30, 2020 as PCT Publication No. WO 2020/261156 A1, including, forexample, a compound selected from:

-   N-(3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-2,4-difluorophenyl)propane-1-sulfonamide;-   N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-3-fluoropropane-1-sulfonamide;-   N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4,5-difluorophenyl)propane-1-sulfonamide;-   N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)propane-1-sulfonamide;-   N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoropropane-1-sulfonamide;-   N-(2-chloro-4-fluoro-3-((5-methyl-3-(methyl-d3)-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-phenyl)-3-fluoropropane-1-sulfonamide;-   N-{2-chloro-3-[(3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy]-4-fluorophenyl}propane-1-sulfonamide;-   N-(3-chloro-4-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy)-5-fluoropyridin-2-yl)propane-1-sulfonamide;    and-   N-{2-chloro-3-[(3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy]-4-fluorophenyl}-3-fluoropropane-1-sulfonamide;    -   or a pharmaceutically acceptable salt thereof;    -   and the compounds disclosed in PCT Publication No. WO        2021/250521, published Dec. 16, 2021, including, for example,        N-(2-chloro-4-fluoro-3-((5-fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-2-azabicyclo[2.1.1]hexane-2-sulfonamide,        (R)-N-(2-chloro-4-fluoro-3-((5-fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-3-fluoropyrrolidine-1-sulfonamide,        and        N-(2-chloro-3-((5-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoroazetidine-1-sulfonamide,        or a pharmaceutically acceptable salt thereof.

In one embodiment, the BRAF inhibitor is selected from encorafenib or apharmaceutically acceptable salt thereof,N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoropropane-1-sulfonamideor a pharmaceutically acceptable salt thereof, andN-(2-chloro-3-((5-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoroazetidine-1-sulfonamideor a pharmaceutically acceptable salt thereof.

In one embodiment, the BRAF inhibitor is encorafenib or apharmaceutically acceptable salt thereof. In one embodiment, the BRAFinhibitor isN-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoropropane-1-sulfonamideor a pharmaceutically acceptable salt thereof. In one embodiment, theBRAF inhibitor isN-(2-chloro-3-((5-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoroazetidine-1-sulfonamideor a pharmaceutically acceptable salt thereof.

Additional examples of BRAF inhibitors are known in the art.

In one embodiment, the anticancer agent that can be used in combinationwith a compound of Formula I or a pharmaceutically acceptable saltthereof or a compound of Formula II or a pharmaceutically acceptablesalt thereof according to any of the combination therapy methodsdescribed herein is an EGFR inhibitor. Non-limiting examples of EGFRinhibitors include cetuximab (Erbitux®), panitumumab (Vectibix®),osimertinib (merelectinib, Tagrisso®), erlotinib (Tarceva®), gefitinib(Iressa®), necitumumab (Portrazza™), neratinib (Nerlynx®), lapatinib(Tykerb®), vandetanib (Caprelsa®), brigatinib (Alunbrig®) and inhibitorsof EGFR disclosed in PCT Publication Nos. WO 2019/071351 and WO2017/117680. Additional examples of EGFR inhibitors are known in theart.

In one embodiment, the anticancer agent that can be used in combinationwith a compound of Formula I or a pharmaceutically acceptable saltthereof or a compound of Formula II or a pharmaceutically acceptablesalt thereof according to any of the combination therapy methodsdisclosed herein is a SHP2 inhibitor. Non-limiting examples of SHP2inhibitors include6-(4-amino-4-methylpiperidin-1-yl)-3-(2,3-dichlorophenyl)pyrazin-2-amine(SHP099),[3-[(3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl]-6-(2,3-dichlorophenyl)-5-methylpyrazin-2-yl]methanol(RMC-4550) RMC-4630, TNO155, and the compounds disclosed in WO2020/081848, WO 2020/201991, WO 2015/107493, WO 2015/107494, WO2015/107495 and WO 2019/075265. In one embodiment, the SHP2 inhibitor isa compound disclosed in WO 2020/201991. In one embodiment, the SHP2inhibitor is(S)-1′-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4′-piperidin]-1-amineor a pharmaceutically acceptable salt thereof.

In one embodiment, the anticancer agent that can be used in combinationwith a compound of Formula I or a pharmaceutically acceptable saltthereof or a compound of Formula II or a pharmaceutically acceptablesalt thereof according to any of the combination therapy methodsdisclosed herein is a PI3K inhibitor. Non-limiting examples includebuparlisib (BKM120), alpelisib (BYL719), samotolisib (LY3023414),8-[(1R)-1-[(3,5-difluorophenyl)amino]ethyl]-N,N-dimethyl-2-(morpholin-4-yl)-4-oxo-4H-chromene-6-carboxamide(AZD8186), tenalisib (RP6530), voxtalisib hydrochloride (SAR-245409),gedatolisib (PF-05212384), panulisib (P-7170), taselisib (GDC-0032),trans-2-amino-8-[4-(2-hydroxyethoxy)cyclohexyl]-6-(6-methoxypyridin-3-yl)-4-methylpyrido[2,3-d]pyrimidin-7(8H)-one(PF-04691502), duvelisib (ABBV-954),N2-[4-oxo-4-[4-(4-oxo-8-phenyl-4H-1-benzopyran-2-yl)morpholin-4-ium-4-ylmethoxy]butyryl]-L-arginyl-glycyl-L-aspartyl-L-serineacetate (SF-1126), pictilisib (GDC-0941),2-methyl-1-[2-methyl-3-(trifluoromethyl)benzyl]-6-(morpholin-4-yl)-1H-benzimidazole-4-carboxylicacid (GSK2636771), idelalisib (GS-1101), umbralisib tosylate (TGR-1202),pictilisib (GDC-0941), copanlisib hydrochloride (BAY 84-1236),dactolisib (BEZ-235),1-(4-[5-[5-amino-6-(5-tert-butyl-1,3,4-oxadiazol-2-yl)pyrazin-2-yl]-1-ethyl-1H-1,2,4-triazol-3-yl]piperidin-1-yl)-3-hydroxypropan-1-one(AZD-8835),5-[6,6-dimethyl-4-(morpholin-4-yl)-8,9-dihydro-6H-[1,4]oxazino[4,3-e]purin-2-yl]pyrimidin-2-amine(GDC-0084) everolimus, rapamycin, perifosine, sirolimus, andtemsirolimus.

In one embodiment, the anticancer agent that can be used in combinationwith a compound of Formula I or a pharmaceutically acceptable saltthereof or a compound of Formula II or a pharmaceutically acceptablesalt thereof according to any of the combination therapy methodsdisclosed herein is an immunotherapy. The term “immunotherapy” refers toan agent that modulates the immune system. In one embodiment, animmunotherapy can increase the expression and/or activity of a regulatorof the immune system. In one embodiment, an immunotherapy can decreasethe expression and/or activity of a regulator of the immune system. Inone embodiment, an immunotherapy can recruit and/or enhance the activityof an immune cell.

In one embodiment, the immunotherapy that can be used in combinationwith a compound of Formula I or a pharmaceutically acceptable saltthereof or a compound of Formula II or a pharmaceutically acceptablesalt thereof according to any of the combination therapy methodsdisclosed herein is an antibody therapy (e.g., a monoclonal antibody, aconjugated antibody). In one embodiment, the antibody therapy isbevacizumab (Mvasti™, Avastin®), trastuzumab (Herceptin®), rituximab(MabThera™, Rituxan®), edrecolomab (Panorex), daratumuab (Darzalex®),olaratumab (Lartruvo™), ofatumumab (Arzerra®), alemtuzumab (Campath®),cetuximab (Erbitux®), oregovomab, pembrolizumab (Keytruda®), dinutiximab(Unituxin®), obinutuzumab (Gazyva®), tremelimumab (CP-675,206),ramucirumab (Cyramza®), ublituximab (TG-1101), panitumumab (Vectibix®),elotuzumab (Empliciti™), necitumumab (Portrazza™), cirmtuzumab (UC-961),ibritumomab (Zevalin®), isatuximab (SAR650984), nimotuzumab,fresolimumab (GC1008), lirilumab (INN), mogamulizumab (Poteligeo®),ficlatuzumab (AV-299), denosumab (Xgeva®), ganitumab, urelumab,pidilizumab, amatuximab, blinatumomab (AMG103; Blincyto®) or midostaurin(Rydapt).

In one embodiment, the immunotherapy that can be used in combinationwith a compound of Formula I or a pharmaceutically acceptable saltthereof or a compound of Formula II or a pharmaceutically acceptablesalt thereof according to any of the combination therapy methodsdisclosed herein is an immune checkpoint inhibitor. In one embodiment,the immunotherapy includes one or more, e.g., one or two immunecheckpoint inhibitors. In one embodiment, the immune checkpointinhibitor is a CTLA-4 inhibitor, a PD-1 inhibitor or a PD-L1 inhibitor.In one embodiment, the CTLA-4 inhibitor is ipilimumab (Yervoy®) ortremelimumab (CP-675,206). In one embodiment, the PD-1 inhibitor ispembrolizumab (Keytruda®), nivolumab (Opdivo®) and sasanlimab (RN888).In one embodiment, the PD-L1 inhibitor is atezolizumab (Tecentriq®), ordurvalumab (Imfinzi™).

In one embodiment, the anticancer therapy that can be used incombination with a compound of Formula I or a pharmaceuticallyacceptable salt thereof or a compound of Formula II or apharmaceutically acceptable salt thereof according to any of thecombination therapy methods disclosed herein is radiotherapy.Non-limiting examples of radiotherapy include external radiation beamtherapy (e.g., external beam therapy using kilovoltage X-rays ormegavoltage X-rays) or internal radiotherapy. Internal radiotherapy(also called brachytherapy) can include the use of, e.g., low-doseinternal radiotherapy or high-dose internal radiotherapy. Low-doseinternal radiotherapy includes, e.g., inserting small radioactivepellets into or proximal to a cancer tissue in the subject. High-doseinternal radiotherapy includes, e.g., inserting a thin tube (e.g., acatheter) or an implant into or proximal to a cancer tissue in thesubject, and delivering a high dose of radiation to the thin tube orimplant using a radiation machine. Methods for performing radiotherapyon a subject having a cancer are known in the art. In embodimentswherein the tumor is a CNS tumor, the radiotherapy may include wholebrain radiotherapy (WBRT) or stereotactic radiosurgery (SRS) such asCyberknife®, XKnife®, Gamma Knife®, or ExacTrac®.

In one embodiment, the anticancer therapy that can be used incombination with a compound of Formula I or a pharmaceuticallyacceptable salt thereof or a compound of Formula II or apharmaceutically acceptable salt thereof according to any of thecombination therapy methods disclosed herein is surgery. Non-limitingexamples of surgery include, e.g., open surgery or minimally invasivesurgery. Surgery can include, e.g., at least a partial resection of thetumor, removing an entire tumor, debulking of a tumor, or removing atumor that is causing pain or pressure in the subject. Methods forperforming open surgery and minimally invasive surgery on a subjecthaving a cancer are known in the art.

In one embodiment, provided herein is a method of treating aMEK-associated tumor (e.g., any of the MEK-associated tumors describedherein) comprising administering to a subject in need thereof atherapeutically effective amounts of a compound of Formula I or apharmaceutically acceptable salt thereof or a compound of Formula II ora pharmaceutically acceptable salt thereof and a BRAF inhibitor (e.g.,any of the BRAF inhibitors disclosed herein) in any order, together orseparately. In one embodiment, the compound of Formula I is a compoundselected from Examples 1-69, or a pharmaceutically acceptable saltthereof.

In one embodiment, provided herein is a method of treating aMEK-associated tumor (e.g., any of the MEK-associated tumors describedherein) comprising administering to a subject in need thereof atherapeutically effective amounts of a compound of Formula I or apharmaceutically acceptable salt thereof or a compound of Formula II ora pharmaceutically acceptable salt thereof and an EGFR inhibitor (e.g.,any of the EGFR inhibitors disclosed herein) in any order, together orseparately. In one embodiment, the compound of Formula I is a compoundselected from any one of Examples 1-69, or a pharmaceutically acceptablesalt thereof.

In one embodiment, provided herein is a method of treating aMEK-associated tumor (e.g., any of the MEK-associated tumors describedherein) comprising administering to a subject in need thereof atherapeutically effective amounts of a compound of Formula I or apharmaceutically acceptable salt thereof or a compound of Formula II ora pharmaceutically acceptable salt thereof and a SHP2 inhibitor (e.g.,any of the SHP2 inhibitors disclosed herein) in any order, together orseparately. In one embodiment, the compound of Formula I is a compoundselected from any one of Examples 1-69, or a pharmaceutically acceptablesalt thereof.

In one embodiment, provided herein is a method of treating aMEK-associated tumor (e.g., any of the MEK-associated tumors describedherein) comprising administering to a subject in need thereof atherapeutically effective amounts of a compound of Formula I or apharmaceutically acceptable salt thereof or a compound of Formula II ora pharmaceutically acceptable salt thereof and a checkpoint inhibitor(e.g., any of the checkpoint inhibitors disclosed herein) in any order,together or separately. In one embodiment, the compound of Formula I isa compound selected from any one of Examples 1-69, or a pharmaceuticallyacceptable salt thereof.

Also provided herein is a pharmaceutical combination for treating aMEK-associated tumor in a subject in need thereof, which comprises (a) acompound of Formula I or a pharmaceutically acceptable salt thereof or acompound of Formula II or a pharmaceutically acceptable salt thereof and(b) at least one additional anticancer (e.g., any of the exemplaryadditional anticancer agents described herein or known in the art),wherein the compound of Formula I or a pharmaceutically acceptable saltthereof or the compound of Formula II or a pharmaceutically acceptablesalt thereof and the one or more, e.g., one or more additionalanticancer agents are formulated separately for simultaneous or separateuse for the treatment of the tumor, wherein the amounts of the compoundof Formula I or a pharmaceutically acceptable salt thereof or thecompound of Formula II or a pharmaceutically acceptable salt thereof andof the additional anticancer agent(s) are together effective in treatingthe tumor; (ii) the use of such a combination for the preparation of amedicament for the treatment of the tumor; and (iii) a commercialpackage or product comprising such a combination as a combinedpreparation for simultaneous, separate or sequential use; and to amethod of treatment of a tumor in a subject in need thereof.

The term “pharmaceutical combination”, as used herein, refers to anon-fixed combination of the active ingredients. The term “non-fixedcombination” means that a compound of Formula I or a pharmaceuticallyacceptable salt thereof or a compound of Formula II or apharmaceutically acceptable salt thereof and one or more, e.g., one ormore additional anticancer agents are formulated as separatecompositions or dosages such that they may be administered to a subjectin need thereof simultaneously or separately with variable interveningtime limits and in any order, wherein such administration provideseffective levels of the two or more compounds in the body of thesubject. These also apply to cocktail therapies, e.g. the administrationof three or more active ingredients. Similarly, the term “combination”when referring to a compound of Formula I or a pharmaceuticallyacceptable salt thereof or a compound of Formula II or apharmaceutically acceptable salt thereof in use with a combination ofone or more anticancer agents refers to a non-fixed combination.

Accordingly, also provided herein is a method of treating aMEK-associated tumor, comprising administering to a subject in needthereof a pharmaceutical combination for treating said tumor whichcomprises (a) a compound of Formula I or a pharmaceutically acceptablesalt thereof or a compound of Formula II or a pharmaceuticallyacceptable salt thereof and (b) one or more, e.g., one or moreadditional anticancer agents for simultaneous, separate or sequentialuse for the treatment of the tumor, wherein the amounts of the compoundof Formula I or a pharmaceutically acceptable salt thereof or thecompound of Formula II or a pharmaceutically acceptable salt thereof andthe additional anticancer agent(s) are together effective in treatingthe tumor.

In one embodiment, provided herein is a method of treating aMEK-associated tumor (e.g., a benign, malignant, or metastatic tumor)comprising administering to a subject in need thereof a therapeuticallyeffective amount of a compound of Formula I or a pharmaceuticallyacceptable salt thereof or a compound of Formula II or apharmaceutically acceptable salt thereof, wherein the subject has notreceived treatment with an anticancer therapy prior to administration ofsaid compound of Formula I or a pharmaceutically acceptable salt thereofor a compound of Formula II or a pharmaceutically acceptable saltthereof, wherein the anticancer therapy is selected from one or more,e.g., one or more anticancer therapies independently selected fromsurgery, radiotherapy and an anticancer agent that works by the same ordifferent mechanism of action. In one embodiment, patient has not beentreated with an anticancer agent prior to administration of saidcompound of Formula I or a pharmaceutically acceptable salt thereof or acompound of Formula II or a pharmaceutically acceptable salt thereof. Inone embodiment, patient has not been treated with surgery prior toadministration of said compound of Formula I or a pharmaceuticallyacceptable salt thereof or a compound of Formula II or apharmaceutically acceptable salt thereof. In one embodiment, patient hasnot been treated with radiotherapy prior to administration of saidcompound of Formula I or a pharmaceutically acceptable salt thereof or acompound of Formula II or a pharmaceutically acceptable salt thereof.

In one embodiment, provided herein is a method of treating a subjecthaving a MEK-associated tumor (e.g., a benign, malignant, or metastatictumor) comprising administering a therapeutically effective amount of acompound of Formula I or a pharmaceutically acceptable salt thereof or acompound of Formula II or a pharmaceutically acceptable salt thereof,wherein the subject has been treated with prior therapy or standardtherapy (e.g., treatment with one or more anticancer agents other than acompound of Formula I or a pharmaceutically acceptable salt thereofand/or radiotherapy and/or surgery) wherein the MEK-associated tumor hasbecome refractory or intolerant to said prior therapy. In oneembodiment, the subject developed brain metastasis during said priortreatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, a subject having metastatic melanoma(e.g., a metastatic melanoma having a BRAF V600 mutation or a BRAFfusion) has received treatment with a BRAF inhibitor prior to treatmentwith a compound of Formula I, or a pharmaceutically acceptable saltthereof, or a compound of Formula II, or a pharmaceutically acceptablesalt thereof. In one embodiment, the subject was previously treated witha BRAF inhibitor selected from encorafenib, dabrafenib, vemurafenib,N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide,(3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide(PLX8394). In one embodiment, the subject was previously treated with aBRAF inhibitor selected from encorafenib, dabrafenib and vemurafenib. Inone embodiment, the subject became refractory to said prior treatment.In one embodiment, the subject developed brain metastasis during saidprior treatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, a subject having metastatic melanoma(e.g., metastatic melanoma having a BRAF V600 mutation or a BRAF fusion)has received treatment with a BRAF inhibitor and a MEK inhibitor priorto treatment with a compound of Formula I, or a pharmaceuticallyacceptable salt thereof, or a compound of Formula II, or apharmaceutically acceptable salt thereof. In one embodiment, the subjectwas previously treated with a BRAF inhibitor selected from encorafenib,dabrafenib, vemurafenib,N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide,and(3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide(PLX8394), and a MEK inhibitor selected from binimetinib, trametinib,cobimetinib, selumetinib, pimasertib, refametinib,N-[2(R),3-dihydroxypropoxy]-3,4-difluoro-2-(2-fluoro-4-iodophenylamino)benzamide(PD-325901),2-(2-chloro-4-iodophenylamino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide(CI-1040), and3-[2(R),3-dihydroxypropyl]-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione(TAK-733). In one embodiment, the subject was previously treated with aBRAF inhibitor selected from encorafenib, dabrafenib and vemurafenib anda MEK inhibitor selected from binimetinib, trametinib, and cobimetinib.In one embodiment, the subject was previously treated with encorafeniband binimetinib. In one embodiment, the subject was previously treatedwith dabrafenib and trametinib. In one embodiment, the subject waspreviously treated with vemurafenib and cobimetinib. In one embodiment,the subject became refractory to said prior treatment. In oneembodiment, the subject developed brain metastasis during said priortreatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, a subject having metastatic melanoma(e.g., metastatic melanoma having a BRAF V600 mutation or a BRAF fusion)has received treatment with one or more, e.g., one or two checkpointinhibitors (e.g., any of the checkpoint inhibitors disclosed herein,e.g., a CTLA-4 inhibitor, a PD-1 inhibitor, and/or a PD-L1 inhibitor)prior to treatment with a compound of Formula I, or a pharmaceuticallyacceptable salt thereof. In one embodiment, the subject was previouslytreated with one or more, e.g., one or two checkpoint inhibitorsindependently selected from ipilimumab, nivolumab, and pembrolizumab. Inone embodiment, the subject became refractory to said prior treatment.In one embodiment, the subject developed brain metastasis during saidprior treatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, a subject having metastatic melanoma(e.g., metastatic melanoma having a BRAF V600 mutation or a BRAF fusion)has received treatment with one or more, e.g., one or two inhibitors ofPI3K prior to treatment with a compound of Formula I, or apharmaceutically acceptable salt thereof, or a compound of Formula II,or a pharmaceutically acceptable salt thereof. In one embodiment, thesubject was previously treated with one or more, e.g., one or two PI3Kinhibitors selected from buparlisib (BKM120), alpelisib (BYL719),samotolisib (LY3023414),8-[(1R)-1-[(3,5-difluorophenyl)amino]ethyl]-N,N-dimethyl-2-(morpholin-4-yl)-4-oxo-4H-chromene-6-carboxamide(AZD8186), tenalisib (RP6530), voxtalisib hydrochloride (SAR-245409),gedatolisib (PF-05212384), panulisib (P-7170), taselisib (GDC-0032),trans-2-amino-8-[4-(2-hydroxyethoxy)cyclohexyl]-6-(6-methoxypyridin-3-yl)-4-methylpyrido[2,3-d]pyrimidin-7(8H)-one(PF-04691502), duvelisib (ABBV-954),N2-[4-oxo-4-[4-(4-oxo-8-phenyl-4H-1-benzopyran-2-yl)morpholin-4-ium-4-ylmethoxy]butyryl]-L-arginyl-glycyl-L-aspartyl-L-serineacetate (SF-1126), pictilisib (GDC-0941),2-methyl-1-[2-methyl-3-(trifluoromethyl)benzyl]-6-(morpholin-4-yl)-1H-benzimidazole-4-carboxylicacid (GSK2636771), idelalisib (GS-1101), umbralisib tosylate (TGR-1202),pictilisib (GDC-0941), copanlisib hydrochloride (BAY 84-1236),dactolisib (BEZ-235),1-(4-[5-[5-Amino-6-(5-tert-butyl-1,3,4-oxadiazol-2-yl)pysrazin-2-yl]-1-ethyl-1H-1,2,4-triazol-3-yl]piperidin-1-yl)-3-hydroxypropan-1-one(AZD-8835),5-[6,6-Dimethyl-4-(morpholin-4-yl)-8,9-dihydro-6H-[1,4]oxazino[4,3-e]purin-2-yl]pyrimidin-2-amine(GDC-0084) everolimus, rapamycin, perifosine, sirolimus, andtemsirolimus. In one embodiment, the subject was previously treated withbuparlisib or alpelisib, alone or in combination. In one embodiment, thesubject became refractory to said prior treatment. In one embodiment,the subject developed brain metastasis during said prior treatment. Inone embodiment, the subject developed brain metastasis during said priortreatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, a subject having metastatic melanoma(e.g., metastatic melanoma having a BRAF V600 mutation or a BRAF fusion)has received treatment with a BRAF inhibitor and one or more, e.g., oneor two checkpoint inhibitors (e.g., any of the checkpoint inhibitorsdisclosed herein, e.g., a CTLA-4 inhibitor, a PD-1 inhibitor, and/or aPD-L1 inhibitor) prior to treatment with a compound of Formula I, or apharmaceutically acceptable salt thereof, or a compound of Formula II,or a pharmaceutically acceptable salt thereof. In one embodiment, thesubject was previously treated with a BRAF inhibitor selected fromencorafenib, dabrafenib, vemurafenib,N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide,and(3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide(PLX8394), and one or more, e.g., one or two checkpoint inhibitorsindependently selected from ipilimumab, nivolumab and pembrolizumab. Inone embodiment, the subject became refractory to said prior treatment.In one embodiment, the subject developed brain metastasis during saidprior treatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, a subject having metastatic melanoma(e.g., metastatic melanoma having a BRAF V600 mutation or a BRAF fusion)has received treatment with a BRAF inhibitor, a MEK inhibitor, and oneor more, e.g., one or two checkpoint inhibitors (e.g., any of thecheckpoint inhibitors disclosed herein, e.g., a CTLA-4 inhibitor, a PD-1inhibitor, and/or a PD-L1 inhibitor) prior to treatment with a compoundof Formula I, or a pharmaceutically acceptable salt thereof, or acompound of Formula II, or a pharmaceutically acceptable salt thereof.In one embodiment, the subject was previously treated with a BRAFinhibitor selected from encorafenib, dabrafenib, vemurafenib,N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide(PLX4720), and(3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide(PLX8394), a MEK inhibitor selected from binimetinib, trametinib,cobimetinib, selumetinib, pimasertib, refametinib,N-[2(R),3-dihydroxypropoxy]-3,4-difluoro-2-(2-fluoro-4-iodophenylamino)benzamide(PD-325901),2-(2-chloro-4-iodophenylamino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide(CI-1040), and3-[2(R),3-dihydroxypropyl]-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione(TAK-733), and one or checkpoint inhibitors (e.g., any of the checkpointinhibitors disclosed herein, e.g., a CTLA-4 inhibitor, a PD-1 inhibitor,and/or a PD-L1 inhibitor). In one embodiment, the subject was previouslytreated with a BRAF inhibitor selected from encorafenib, dabrafenib andvemurafenib, a MEK inhibitor selected from binimetinib, trametinib andcobimetinib, and one or more, e.g., one or two checkpoint inhibitorsindependently selected from ipilimumab, nivolumab, and pembrolizumab. Inone embodiment, the subject became refractory to said prior treatment.In one embodiment, the subject developed brain metastasis during saidprior treatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, a subject having metastatic melanoma(e.g., metastatic melanoma having a BRAF V600 mutation or a BRAF fusion)has received treatment with one or more, e.g., one or two alkylatingagents prior to treatment with a compound of Formula I, or apharmaceutically acceptable salt thereof, or a compound of Formula II,or a pharmaceutically acceptable salt thereof. In one embodiment, thesubject was previously treated with s alkylating agents selected fromtemozolomide, fotemustine, lomustine and carmustine. In one embodiment,the subject was previously treated with temozolomide. In one embodiment,the subject became refractory to said prior treatment. In oneembodiment, the subject developed brain metastasis during said priortreatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, a subject having metastatic colorectalcancer (e.g., a metastatic colorectal cancer having a BRAF V600 mutationor a BRAF fusion) has received treatment with a MEK inhibitor and one ormore, e.g., one or two checkpoint inhibitors (e.g., any of thecheckpoint inhibitors disclosed herein, e.g., a CTLA-4 inhibitor, a PD-1inhibitor, and/or a PD-L1 inhibitor) prior to treatment with a compoundof Formula I, or a pharmaceutically acceptable salt thereof, or FormulaII or a pharmaceutically acceptable salt thereof. In one embodiment, thesubject was previously treated with a MEK inhibitor selected frombinimetinib, trametinib, cobimetinib, selumetinib, pimasertib,refametinib,N-[2(R),3-dihydroxypropoxy]-3,4-difluoro-2-(2-fluoro-4-iodophenylamino)benzamide(PD-325901),2-(2-chloro-4-iodophenylamino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide(CI-1040), and3-[2(R),3-dihydroxypropyl]-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione(TAK-733), and one or checkpoint inhibitors (e.g., any of the checkpointinhibitors disclosed herein, e.g., a CTLA-4 inhibitor, a PD-1 inhibitor,and/or a PD-L1 inhibitor). In one embodiment, the subject was previouslytreated with a MEK inhibitor selected from binimetinib, trametinib andcobimetinib, and one or more, e.g., one or two checkpoint inhibitorsindependently selected from ipilimumab, nivolumab, and pembrolizumab. Inone embodiment, the subject was previously treated with a MEK inhibitorwhich is binimetinib and a checkpoint inhibitor which is nivolumab,ipilimumab, or pembrolizumab. In one embodiment, the subject becamerefractory to said prior treatment. In one embodiment, the subjectdeveloped brain metastasis during said prior treatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, a subject having metastatic colorectalcancer (e.g., a metastatic colorectal cancer having a BRAF V600 mutationor a BRAF fusion) has received treatment with one or more, e.g., one ortwo checkpoint inhibitors (e.g., any of the checkpoint inhibitorsdisclosed herein, e.g., a CTLA-4 inhibitor, a PD-1 inhibitor, and/or aPD-L1 inhibitor) prior to treatment with a compound of Formula I, or apharmaceutically acceptable salt thereof, or a compound of Formula II,or a pharmaceutically acceptable salt thereof. In one embodiment, thesubject was previously treated with one or more, e.g., one or twocheckpoint inhibitors independently selected from ipilimumab, nivolumab,pembrolizumab, and sasanlimab. In one embodiment, the subject becamerefractory to said prior treatment. In one embodiment, the subjectdeveloped brain metastasis during said prior treatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, a subject having a metastatic colorectalcancer (e.g., mutant metastatic colorectal cancer having a BRAF V600mutation or a BRAF fusion) has received treatment with one or morecytotoxic chemotherapeutic agents prior to treatment with a compound ofFormula I, or a pharmaceutically acceptable salt, thereof, or a compoundof Formula II, or a pharmaceutically acceptable salt thereof. In oneembodiment, the subject has received treatment with oxaliplatin,irinotecan, FOLFOXIRI (oxaliplatin, irinotecan and fluorouracil),FOLFIRI (folinic acid, fluorouracil and irinotecan) or CAPEOX(capecitabine and oxaliplatin) prior to treatment with a compound ofFormula I, or a pharmaceutically acceptable salt, thereof. In oneembodiment, the subject became refractory to said prior treatment. Inone embodiment, the subject developed brain metastasis during said priortreatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, a subject having a metastatic colorectalcancer (e.g., metastatic colorectal cancer having a BRAF V600 mutationor a BRAF fusion) has received treatment with an EGFR inhibitor, a BRAFinhibitor, and one or more cytotoxic chemotherapeutic agents prior totreatment with a compound of Formula I, or a pharmaceutically acceptablesalt thereof, or a compound of Formula II, or a pharmaceuticallyacceptable salt thereof. In one embodiment, a subject having ametastatic colorectal cancer previously received treatment with an EGFRinhibitor selected from cetuximab, panitumumab, osimertinib, erlotinib,gefitinib, necitumumab, neratinib, lapatinib, vandetanib and brigatinib,a BRAF inhibitor selected from encorafenib, dabrafenib, vemurafenib,N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide(PLX4720), and(3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide(PLX8394), and one or more cytotoxic chemotherapeutic agents. In oneembodiment, the subject previously received treatment with an EGFRinhibitor selected from cetuximab, and panitumumab, a BRAF inhibitorwhich is vemurafenib, and a cytotoxic chemotherapeutic agent which isirinotecan. In one embodiment, the subject became refractory to saidprior treatment. In one embodiment, the subject developed brainmetastasis during said prior treatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, a subject having a metastatic colorectalcancer (e.g., a metastatic colorectal cancer having a BRAF V600 mutationor a BRAF fusion) has received treatment with an EGFR inhibitor and oneor more cytotoxic chemotherapeutic agents prior to treatment with acompound of Formula I, or a pharmaceutically acceptable salt thereof, acompound of Formula II, or a pharmaceutically acceptable salt thereof.In one embodiment, the subject has previously received treatment with anEGFR inhibitor selected from cetuximab, panitumumab, osimertinib,erlotinib, gefitinib, necitumumab, neratinib, lapatinib, vandetanib andbrigatinib, and one or more chemotherapeutic agents. In one embodiment,the subject has previously received treatment with an EGFR inhibitorselected from cetuximab, and panitumumab, and a cytotoxicchemotherapeutic agent which is irinotecan or FOLFIRI (folinic acid,fluorouracil and irinotecan). In one embodiment, the subject becamerefractory to said prior treatment. In one embodiment, the subjectdeveloped brain metastasis during said prior treatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, a subject having metastatic non-smallcell lung cancer (e.g., a metastatic non-small cell lung cancer having aBRAF V600 mutation or a BRAF fusion) has received treatment with one ormore, e.g., one or two EGFR inhibitors prior to treatment with acompound of Formula I, or a pharmaceutically acceptable salt thereof, acompound of Formula II, or a pharmaceutically acceptable salt thereof.In one embodiment, the subject was previously treated with one or more,e.g., one or two EGFR inhibitors independently selected from cetuximab,panitumumab, osimertinib, erlotinib, gefitinib, necitumumab, neratinib,lapatinib, vandetanib and brigatinib. In one embodiment, the subject waspreviously treated with erlotinib. In one embodiment, the subject waspreviously treated with gefitinib. In one embodiment, the subject waspreviously treated with erlotinib and gefitinib. In one embodiment, thesubject became refractory to said prior treatment. In one embodiment,the subject developed brain metastasis during said prior treatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, a subject having metastatic non-smallcell lung cancer (e.g., metastatic non-small cell lung cancer having aBRAF mutation) previously received treatment with a BRAF inhibitorselected from encorafenib, dabrafenib, vemurafenib,N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide(PLX4720), and(3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide(PLX8394), a MEK inhibitor selected from binimetinib, trametinib,cobimetinib, selumetinib, pimasertib, refametinib,N-[2(R),3-dihydroxypropoxy]-3,4-difluoro-2-(2-fluoro-4-iodophenylamino)benzamide(PD-325901),2-(2-chloro-4-iodophenylamino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide(CI-1040), and3-[2(R),3-dihydroxypropyl]-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione(TAK-733), and an EGFR inhibitor selected from cetuximab, panitumumab,osimertinib, erlotinib, gefitinib, necitumumab, neratinib, lapatinib,vandetanib and brigatinib prior to treatment with a compound of FormulaI or a pharmaceutically acceptable salt thereof or a compound of FormulaII or a pharmaceutically acceptable salt thereof. In one embodiment, thesubject was previously treated with a BRAF inhibitor selected fromvemurafenib, dabrafenib and encorafenib and an EGFR inhibitor selectedfrom cetuximab and panitumumab prior to treatment with a compound ofFormula I, or a pharmaceutically acceptable salt thereof. In oneembodiment, the subject became refractory to said prior treatment. Inone embodiment, the subject developed brain metastasis during said priortreatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, a subject having metastatic thyroidcancer (e.g., metastatic thyroid cancer having a BRAF V600 mutation or aBRAF fusion) previously received treatment with a BRAF inhibitorselected from encorafenib, dabrafenib, vemurafenib,N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide(PLX4720), and(3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide(PLX8394), a MEK inhibitor selected from binimetinib, trametinib,cobimetinib, selumetinib, pimasertib, refametinib,N-[2(R),3-dihydroxypropoxy]-3,4-difluoro-2-(2-fluoro-4-iodophenylamino)benzamide(PD-325901),2-(2-chloro-4-iodophenylamino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide(CI-1040), and3-[2(R),3-dihydroxypropyl]-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione(TAK-733), and an EGFR inhibitor selected from cetuximab, panitumumab,osimertinib, erlotinib, gefitinib, necitumumab, neratinib, lapatinib,vandetanib and brigatinib prior to treatment with a compound of FormulaI, or a pharmaceutically acceptable salt thereof, a compound of FormulaII, or a pharmaceutically acceptable salt thereof. In one embodiment,the subject was previously treated with a BRAF inhibitor selected fromvemurafenib, dabrafenib and encorafenib prior to treatment with acompound of Formula I, or a pharmaceutically acceptable salt thereof. Inone embodiment, the subject became refractory to said prior treatment.In one embodiment, the subject developed brain metastasis during saidprior treatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, the subject has LMD and was previouslytreated with a BRAF inhibitor and one or more, e.g., one or twocheckpoint inhibitors (e.g., any of the checkpoint inhibitors disclosedherein, e.g., a CTLA-4 inhibitor, a PD-1 inhibitor, and/or a PD-L1inhibitor) prior to treatment with a compound of Formula I, or apharmaceutically acceptable salt thereof, a compound of Formula II, or apharmaceutically acceptable salt thereof. In one embodiment, the subjectwas previously treated with a BRAF inhibitor selected from encorafenib,dabrafenib, vemurafenib,N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide(PLX4720), and(3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide(PLX8394), and one or more, e.g., one or two checkpoint inhibitorsindependently selected from ipilimumab, nivolumab, and pembrolizumab. Inone embodiment, the subject became refractory to said prior treatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, the subject has LMD and was previouslytreated with a BRAF inhibitor, a MEK inhibitor, and one or more, e.g.,one or two checkpoint inhibitors (e.g., any of the checkpoint inhibitorsdisclosed herein, e.g., a CTLA-4 inhibitor, a PD-1 inhibitor, and/or aPD-L1 inhibitor) prior to treatment with a compound of Formula I, or apharmaceutically acceptable salt thereof, a compound of Formula II, or apharmaceutically acceptable salt thereof. In one embodiment, the subjectwas previously treated with a BRAF inhibitor selected from encorafenib,dabrafenib, vemurafenib,N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide(PLX4720), and(3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide(PLX8394), a MEK inhibitor selected from binimetinib, trametinib,cobimetinib, selumetinib, pimasertib, refametinib,N-[2(R),3-dihydroxypropoxy]-3,4-difluoro-2-(2-fluoro-4-iodophenylamino)benzamide(PD-325901),2-(2-chloro-4-iodophenylamino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide(CI-1040), and3-[2(R),3-dihydroxypropyl]-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione(TAK-733), and a checkpoint inhibitor (e.g., any of the checkpointinhibitors disclosed herein, e.g., a CTLA-4 inhibitor, a PD-1 inhibitor,and/or a PD-L1 inhibitor). In one embodiment, the subject was previouslytreated with a BRAF inhibitor selected from encorafenib, dabrafenib andvemurafenib, a MEK inhibitor selected from binimetinib, trametinib andcobimetinib, and one or more, e.g., one or two checkpoint inhibitorsindependently selected from ipilimumab, nivolumab, and pembrolizumab. Inone embodiment, the subject became refractory to said prior treatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, the subject has LMD and was previouslytreated with one or more, e.g., one or two checkpoint inhibitors (e.g.,any of the checkpoint inhibitors disclosed herein, e.g., a CTLA-4inhibitor, a PD-1 inhibitor, and/or a PD-L1 inhibitor) prior totreatment with a compound of Formula I, or a pharmaceutically acceptablesalt thereof, a compound of Formula II, or a pharmaceutically acceptablesalt thereof. In one embodiment, the subject was previously treated withone or more, e.g., one or two checkpoint inhibitors independentlyselected from ipilimumab, nivolumab, and pembrolizumab. In oneembodiment, the subject became refractory to said prior treatment.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, the subject has a glioma and waspreviously treated with surgery prior to treatment with a compound ofFormula I, or a pharmaceutically acceptable salt thereof, a compound ofFormula II, or a pharmaceutically acceptable salt thereof. In oneembodiment, the subject became refractory to said prior treatment. Inone embodiment, the glioma is a Grade 2, Grade 3 or Grade 4 glioma.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, the subject has a glioma and waspreviously treated with radiotherapy (e.g., whole brain radiotherapy orstereotactic radiosurgery) prior to treatment with a compound of FormulaI, or a pharmaceutically acceptable salt thereof, a compound of FormulaII, or a pharmaceutically acceptable salt thereof. In one embodiment,the subject became refractory to said prior treatment. In oneembodiment, the glioma is a Grade 2, Grade 3 or Grade 4 glioma.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, the subject has a glioma and waspreviously treated with one or more cytotoxic chemotherapy agents priorto treatment with a compound of Formula I, or a pharmaceuticallyacceptable salt thereof, a compound of Formula II, or a pharmaceuticallyacceptable salt thereof. In one embodiment, the subject was previouslytreated with and one or more cytotoxic chemotherapy agents independentlyselected from cisplatin, pemetrexed, vinorelbine and paclitaxel. In oneembodiment, the subject became refractory to said prior treatment. Inone embodiment, the glioma is a Grade 2, Grade 3 or Grade 4 glioma.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, the subject has a MEK-associated gliomaand was previously treated with an ornithine decarboxylase inhibitorprior to treatment with a compound of Formula I, or a pharmaceuticallyacceptable salt thereof, a compound of Formula II, or a pharmaceuticallyacceptable salt thereof. In one embodiment, the subject previouslyreceived treatment with an ornithine decarboxylase inhibitor which iseflornithine (as the racemate, or D or L enantiomer). In one embodiment,the subject became refractory to said prior treatment. In oneembodiment, the glioma is a Grade 2, Grade 3 or Grade 4 glioma.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, the subject has a MEK-associated gliomaand was previously treated with an alkylating agent prior to treatmentwith a compound of Formula I, or a pharmaceutically acceptable saltthereof, or a pharmaceutically acceptable salt thereof, a compound ofFormula II, or a pharmaceutically acceptable salt thereof. In oneembodiment, the subject previously received treatment with an alkylatingagent selected from temozolomide, lomustine, and carmustine. In oneembodiment, the subject became refractory to said prior treatment. Inone embodiment, the glioma is a Grade 2, Grade 3 or Grade 4 glioma.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, the subject has a MEK-associated gliomaand was previously treated with an alkylating agent and an ornithinedecarboxylase inhibitor prior to treatment with a compound of Formula I,or a pharmaceutically acceptable salt thereof, a compound of Formula II,or a pharmaceutically acceptable salt thereof. In one embodiment, thesubject previously received treatment with an alkylating agent selectedfrom temozolomide, lomustine, and carmustine, and an ornithinedecarboxylase inhibitor which is eflornithine (as the racemate, or D orL enantiomer). In one embodiment, the subject became refractory to saidprior treatment. In one embodiment, the glioma is a Grade 2, Grade 3 orGrade 4 glioma.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, the subject has a MEK-associated gliomaand was previously treated with radiotherapy (e.g., whole brainradiotherapy or stereotactic radiosurgery) and an alkylating agent priorto treatment with a compound of Formula I, or a pharmaceuticallyacceptable salt thereof, a compound of Formula II, or a pharmaceuticallyacceptable salt thereof. In one embodiment, the subject previouslyreceived treatment with radiotherapy (e.g., whole brain radiotherapy orstereotactic radiosurgery) and an alkylating agent selected fromtemozolomide, lomustine, and carmustine. In one embodiment, the subjectbecame refractory to said prior treatment. In one embodiment, the gliomais a Grade 2, Grade 3 or Grade 4 glioma.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, the subject has a MEK-associated gliomaand was previously treated with an antibody therapy prior to treatmentwith a compound of Formula I, or a pharmaceutically acceptable saltthereof, a compound of Formula II, or a pharmaceutically acceptable saltthereof. In one embodiment, the subject previously received treatmentwith an antibody therapy which is bevacizumab. In one embodiment, thesubject became refractory to said prior treatment. In one embodiment,the glioma is a Grade 2, Grade 3 or Grade 4 glioma.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, the subject has a MEK-associated gliomaand was previously treated with surgery and radiotherapy prior totreatment with a compound of Formula I, or a pharmaceutically acceptablesalt thereof, a compound of Formula II, or a pharmaceutically acceptablesalt thereof. In one embodiment, the subject became refractory to saidprior treatment. In one embodiment, the glioma is a Grade 2, Grade 3 orGrade 4 glioma.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, the subject has a MEK-associated gliomaand was previously treated with surgery, radiotherapy and an alkylatingagent prior to treatment with a compound of Formula I, or apharmaceutically acceptable salt thereof, a compound of Formula II, or apharmaceutically acceptable salt thereof. In one embodiment, the subjectwas previously treated with surgery, radiotherapy (e.g., whole brainradiotherapy or stereotactic radiosurgery) and an alkylating agentselected from temozolomide, lomustine, and carmustine. In oneembodiment, the subject became refractory to said prior treatment. Inone embodiment, the glioma is a Grade 2, Grade 3 or Grade 4 glioma.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, the subject has a MEK-associated gliomaand was previously treated with a BRAF inhibitor prior to treatment withcompound of Formula II, or a pharmaceutically acceptable salt thereof.In one embodiment, the subject previously received treatment with a BRAFinhibitor selected fromN-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide(PLX4720), vemurafenib, dabrafenib, encorafenib and(3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide(PLX8394). In one embodiment, the subject became refractory to saidprior treatment. In one embodiment, the glioma is a Grade 2, Grade 3 orGrade 4 glioma.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, the subject has a MEK-associated gliomaand was previously treated with a BRAF inhibitor and a MEK inhibitorprior to treatment with compound of Formula I, or a pharmaceuticallyacceptable salt, solvate or polymorph thereof, or Formula II, or apharmaceutically acceptable salt, solvate or polymorph thereof. In oneembodiment, the subject previously received treatment with a BRAFinhibitor selected fromN-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide(PLX4720), vemurafenib, dabrafenib, encorafenib and(3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide(PLX8394) and a MEK inhibitor selected from binimetinib, trametinib,cobimetinib, selumetinib, pimasertib, refametinib,N-[2(R),3-dihydroxypropoxy]-3,4-difluoro-2-(2-fluoro-4-iodophenylamino)benzamide(PD-325901),2-(2-chloro-4-iodophenylamino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide(CI-1040), and3-[2(R),3-dihydroxypropyl]-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione(TAK-733). In one embodiment, the subject was previously treated with aBRAF inhibitor selected from encorafenib, dabrafenib and vemurafenib anda MEK inhibitor selected from binimetinib, trametinib, and cobimetinib.In one embodiment, the subject became refractory to said priortreatment. In one embodiment, the glioma is a Grade 2, Grade 3 or Grade4 glioma.

In one embodiment of a method disclosed herein for of treating a subjecthaving a MEK-associated tumor, the subject has a MEK-associatedbrainstem ganglioglioma and was previously treated with a BRAF inhibitorprior to treatment with a compound of Formula I, or a pharmaceuticallyacceptable salt thereof, a compound of Formula II, or a pharmaceuticallyacceptable salt thereof. In one embodiment, the subject was previouslytreated with a BRAF inhibitor selected from encorafenib, dabrafenib,vemurafenib,N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide(PLX4720), and(3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide(PLX8394). In one embodiment, the subject was previously treated with aBRAF inhibitor selected from encorafenib, dabrafenib and vemurafenib. Inone embodiment, the subject became refractory to said prior treatment.

Although the genetic basis of tumorigenesis may vary between differentcancer types, the cellular and molecular mechanisms required formetastasis appear to be similar for all solid tumor types. During ametastatic cascade, the cancer cells lose growth inhibitory responses,undergo alterations in adhesiveness and produce enzymes that can degradeextracellular matrix components. This leads to detachment of tumor cellsfrom the original tumor, infiltration into the circulation through newlyformed vasculature, migration and extravasation of the tumor cells atfavorable distant sites where they may form colonies. A number of geneshave been identified as being promoters or suppressors of metastasis.

Accordingly, also provided herein are methods for treating, inhibiting,preventing, aiding in the prevention, or decreasing metastasis of aMEK-associated tumor in a subject in need thereof, the method comprisingadministering to the subject a therapeutically effective amount of acompound of Formula I or a pharmaceutically acceptable salt thereof or acompound of Formula II or a pharmaceutically acceptable salt thereof. Inone embodiment, the compound of Formula I or a pharmaceuticallyacceptable salt thereof, is used in combination with one or moreanticancer therapies independently selected from surgery (e.g., at leastpartial resection of a tumor), radiotherapy, and an anticancer agent.

As used herein, the term “treating metastasis” means reducing the size,progression, and/or further spread of one or more metastases.

As used herein, the term “inhibiting metastasis” means reducing theoccurrence (or reoccurrence) of one or more metastases, preventing theoccurrence (or reoccurrence) of one or more metastases, or reducing thespread of one or more metastases.

In one embodiment, a subject treated according to any of the methodsdisclosed herein may be assessed according to one or more standardresponse assessment criteria known in the art, including RECIST(Response Evaluation Criteria in Solid Tumors, e.g., RECIST version 1.0,RECIST version 1.1, and modified RECIST 1.1 (mRECIST 1.1)), RANO-BM(Response Assessment in Neuro-Oncology Brain Metastases), Macdonald,RANO-LMD, and NANO (Neurologic Assessment in Neuro-Oncology). In oneembodiment of any of said criteria, the tumor is assessed by an imagingstudy (e.g., MRI, CT, MDCT or PET). In one embodiment the treatmentresponse is assessed in accordance with RECIST version 1.1, wherein:complete response (CR) is defined as the complete disappearance of alltumor lesions; partial response (PR) is defined as a reduction in thesum of tumor measurements by at least 30%; progressive disease (PD) isdefined as at least 20% increase in the sum of tumor measurements(wherein the development of new lesions or substantial progression ofnon-target lesions is also was defined as PD) wherein an increase of atleast 5 mm from baseline is evaluated as PD; and stable disease (SD) isdefined as neither sufficient shrinkage to qualify for PR nor sufficientincrease to qualify for PD, taking as reference the smallest sumdiameters while on treatment. In one embodiment, assessments includeintracranial response (assessed as per modified RECIST using gadoliniumenhanced MRI), extracranial response, global response rate, diseasecontrol rate (DCR), duration of response (DOR), progression freesurvival (PFS), and overall survival (OS).

As used herein, an “effective dosage” or “effective amount” of drug,compound or pharmaceutical composition is an amount sufficient to affectany one or more beneficial or desired, including biochemical,histological and/or behavioral symptoms, of the disease, itscomplications and intermediate pathological phenotypes presenting duringdevelopment of the disease. For therapeutic use, a “therapeuticallyeffective amount” refers to that amount of a compound being administeredwhich will relieve to some extent one or more of the symptoms of thedisorder being treated. In reference to the treatment of cancer, atherapeutically effective amount refers to that amount which has theeffect of (1) reducing the size of the tumor, (2) inhibiting (that is,slowing to some extent, preferably stopping) tumor metastasis, (3)inhibiting to some extent (that is, slowing to some extent, preferablystopping) tumor growth or tumor invasiveness, (4) relieving to someextent (or, preferably, eliminating) one or more signs or symptomsassociated with the cancer, (5) decreasing the dose of other medicationsrequired to treat the disease, and/or (6) enhancing the effect ofanother medication, and/or (7) delaying the progression of the diseasein a patient.

An effective dosage can be administered in one or more administrations.For the purposes of this invention, an effective dosage of drug,compound, or pharmaceutical composition is an amount sufficient toaccomplish prophylactic or therapeutic treatment either directly orindirectly. As is understood in the clinical context, an effectivedosage of drug, compound or pharmaceutical composition may or may not beachieved in conjunction with another drug, compound or pharmaceuticalcomposition.

A “pharmaceutical composition” refers to a mixture of one or more of thecompounds of the invention, or a pharmaceutically acceptable salt,solvate, hydrate or prodrug thereof as an active ingredient, and atleast one pharmaceutically acceptable carrier or excipient. In oneembodiment, the pharmaceutical composition comprises two or morepharmaceutically acceptable carriers and/or excipients.

In one embodiment, the invention provides a pharmaceutical compositioncomprising a compound of the invention, or a pharmaceutically acceptablesalt thereof, and at least one pharmaceutically acceptable carrier orexcipient. In one embodiment, the pharmaceutical composition comprisestwo or more pharmaceutically acceptable carriers and/or excipients.

Accordingly, in one embodiment, the invention provides a pharmaceuticalcomposition for use in the treatment of abnormal cell growth in asubject in need thereof, which pharmaceutical composition comprises acompound of the invention, or a pharmaceutically acceptable saltthereof, and at least one pharmaceutically acceptable carrier orexcipient.

As used herein, a “pharmaceutically acceptable carrier” refers to acarrier or diluent that does not cause significant irritation to anorganism and does not abrogate the biological activity and properties ofthe administered compound.

The pharmaceutical acceptable carrier may comprise any conventionalpharmaceutical carrier or excipient. The choice of carrier and/orexcipient will to a large extent depend on factors such as theparticular mode of administration, the effect of the carrier orexcipient on solubility and stability, and the nature of the dosageform.

Suitable pharmaceutical carriers include inert diluents or fillers,water and various organic solvents (such as hydrates and solvates). Thepharmaceutical compositions may, if desired, contain additionalingredients such as flavorings, binders, excipients and the like.

The term ‘excipient’ is used herein to describe any ingredient otherthan the compound(s) of the invention. The choice of excipient will to alarge extent depend on factors such as the mode of administration, theeffect of the excipient on solubility and stability, and the nature ofthe dosage form.

As used herein, “excipient” includes any and all solvents, dispersionmedia, coatings, antibacterial and antifungal agents, isotonic andabsorption delaying agents, carriers, diluents and the like that arephysiologically compatible. Examples of excipients include one or moreof water, saline, phosphate buffered saline, dextrose, glycerol, ethanoland the like, as well as combinations thereof, and may include isotonicagents, for example, sugars, sodium chloride, or polyalcohols such asmannitol, or sorbitol in the composition. Examples of excipients alsoinclude various organic solvents (such as hydrates and solvates). Thepharmaceutical compositions may, if desired, contain additionalexcipients such as flavorings, binders/binding agents, lubricatingagents, disintegrants, sweetening or flavoring agents, coloring mattersor dyes, and the like. For example, for oral administration, tabletscontaining various excipients, such as citric acid may be employedtogether with various disintegrants such as starch, alginic acid andcertain complex silicates and with binding agents such as sucrose,gelatin and acacia. Examples, without limitation, of excipients includecalcium carbonate, calcium phosphate, various sugars and types ofstarch, cellulose derivatives, gelatin, vegetable oils and polyethyleneglycols. Thus, for oral administration, tablets containing variousexcipients, such as citric acid may be employed together with variousdisintegrants such as starch, alginic acid and certain complex silicatesand with binding agents such as sucrose, gelatin and acacia. Examples,without limitation, of excipients include calcium carbonate, calciumphosphate, various sugars and types of starch, cellulose derivatives,gelatin, vegetable oils and polyethylene glycols. Additionally,lubricating agents such as magnesium stearate, sodium lauryl sulfate andtalc are often useful for tableting purposes. Solid compositions of asimilar type may also be employed in soft and hard filled gelatincapsules. Non-limiting examples of materials, therefore, include lactoseor milk sugar and high molecular weight polyethylene glycols. Whenaqueous suspensions or elixirs are desired for oral administration theactive compound therein may be combined with various sweetening orflavoring agents, coloring matters or dyes and, if desired, emulsifyingagents or suspending agents, together with diluents such as water,ethanol, propylene glycol, glycerin, or combinations thereof.

Examples of excipients also include pharmaceutically acceptablesubstances such as wetting agents or minor amounts of auxiliarysubstances such as wetting or emulsifying agents, preservatives, orbuffers, which enhance the shelf life or effectiveness of the compound.

The pharmaceutical composition may, for example, be in a form suitablefor oral administration as a tablet, capsule, pill, powder, sustainedrelease formulations, solution suspension, liquid solutions (e.g.,injectable and infusible solutions) for parenteral injection as asterile solution, suspension or emulsion, for topical administration asan ointment or cream, powders, liposomes and suppositories (e.g., forrectal administration as a suppository). Exemplary parenteraladministration forms include solutions or suspensions of activecompounds in sterile aqueous solutions, for example, aqueous propyleneglycol or dextrose solutions. Such dosage forms may be suitablybuffered, if desired. The form depends on the intended mode ofadministration and therapeutic application.

The pharmaceutical composition may be in unit dosage forms suitable forsingle administration of precise dosages.

The compounds of the invention may be administered orally. Oraladministration may involve swallowing, so that the compound enters thegastrointestinal tract, or buccal or sublingual administration may beemployed by which the compound enters the blood stream directly from themouth. Formulations suitable for oral administration include solidformulations such as tablets, capsules containing particulates, liquids,or powders, lozenges (including liquid filled), chews, multi and nanoparticulates, gels, solid solution, liposome, films (including mucoadhesive), ovules, sprays and liquid formulations. Such capsules ortablets may comprise a controlled release formulation. In the case ofcapsules, tablets, and pills, the dosage forms also may comprisebuffering agents or may be prepared with enteric coatings.

Liquid formulations include suspensions, solutions, syrups and elixirs.Such formulations may be used as fillers in soft or hard capsules andtypically include a carrier or adjuvant, for example such as one or moreof wetting, emulsifying, suspending, flavoring (e.g., sweetening), orperfuming agents, for example, water, ethanol, polyethylene glycol,propylene glycol, methylcellulose, or a suitable oil, and one or moreemulsifying agents and/or suspending agents. Liquid formulations mayalso be prepared by the reconstitution of a solid, for example, from asachet.

The compounds of the invention may also be used in fast dissolving, fastdisintegrating dosage forms such as those described in Expert Opinion inTherapeutic Patents, 11 (6), 981 986 by Liang and Chen (2001), thedisclosure of which is incorporated herein by reference in its entirety.

For tablet dosage forms, depending on dose, the drug may make up from 1wt % to 80 wt % of the dosage form, more typically from 5 wt % to 60 wt% of the dosage form. In addition to the drug, tablets generally containa disintegrant. Examples of disintegrants include sodium starchglycolate, sodium carboxymethyl cellulose, calcium carboxymethylcellulose, croscarmellose sodium, crospovidone, polyvinylpyrrolidone,methyl cellulose, microcrystalline cellulose, lower alkyl substitutedhydroxypropyl cellulose, starch, pregelatinized starch and sodiumalginate. Generally, the disintegrant will comprise from 1 wt % to 25 wt%, preferably from 5 wt % to 20 wt % of the dosage form. Binders aregenerally used to impart cohesive qualities to a tablet formulation.Suitable binders include microcrystalline cellulose, gelatin, sugars,polyethylene glycol, natural and synthetic gums, polyvinylpyrrolidone,pregelatinized starch, hydroxypropyl cellulose and hydroxypropylmethylcellulose. Tablets may also contain diluents, such as lactose(monohydrate, spray dried monohydrate, anhydrous and the like),mannitol, xylitol, dextrose, sucrose, sorbitol, microcrystallinecellulose, starch and dibasic calcium phosphate dihydrate. Tablets mayalso optionally include surface active agents, such as sodium laurylsulfate and polysorbate 80, and glidants such as silicon dioxide andtalc. When present, surface active agents are typically in amounts offrom 0.2 wt % to 5 wt % of the tablet, and glidants typically from 0.2wt % to 1 wt % of the tablet. Tablets also generally contain lubricantssuch as magnesium stearate, calcium stearate, zinc stearate, sodiumstearyl fumarate, and mixtures of magnesium stearate with sodium laurylsulphate. Lubricants generally are present in amounts from 0.25 wt % to10 wt %, preferably from 0.5 wt % to 3 wt % of the tablet. Otherconventional ingredients include anti-oxidants, colorants, flavoringagents, preservatives and taste masking agents. Tablet blends may becompressed directly or by roller to form tablets. Tablet blends orportions of blends may alternatively be wet, dry, or melt granulated,melt congealed, or extruded before tableting. The final formulation mayinclude one or more layers and may be coated or uncoated; orencapsulated. Solid formulations for oral administration may beformulated to be immediate and/or modified release. Modified releaseformulations include delayed, sustained pulsed, controlled, targeted andprogrammed release.

For oral administration, the compositions may be provided in the form oftablets or capsules containing 0.01, 0.05, 0.1, 0.25, 0.5, 1.0, 2.5,5.0, 10.0, 15.0, 25.0, 50.0, 75.0, or 100 milligrams of the activeingredient for the symptomatic adjustment of the dosage to the patient.A medicament typically contains from about 0.01 mg to about 100 mg ofthe active ingredient. In another embodiment, a medicament contains fromabout 0.01 to 0.25 mg of the active ingredient. In another embodiment, amedicament contains about 0.25, 0.5, 1.0, 5.0, 15 or 25 mg of the activeingredient.

The compounds of the invention may also be administered directly intothe blood stream, into muscle, or into an internal organ. Suitable meansfor parenteral administration include intravenous, intraarterial,intraperitoneal, intrathecal, intraventricular, intraurethral,intrasternal, intracranial, intramuscular and subcutaneous. Suitabledevices for parenteral administration include needle (including microneedle) injectors, needle free injectors and infusion techniques.Injectable preparations (i.e., sterile injectable aqueous or oleaginoussuspensions) may be formulated according to the known art using one ormore of suitable dispersing, wetting agents, or suspending agents.Parenteral formulations are typically aqueous solutions which maycontain excipients such as salts, carbohydrates and buffering agents(preferably to a pH of from 3 to 9), but, for some applications, theymay be more suitably formulated as a sterile non aqueous solution or asa dried form to be used in conjunction with a suitable vehicle such assterile, pyrogen free water. The preparation of parenteral formulationsunder sterile conditions, for example, by lyophilization, may readily beaccomplished using standard pharmaceutical techniques well known tothose skilled in the art. The solubility of compounds of the inventionused in the preparation of parenteral solutions may be increased by theuse of appropriate formulation techniques, such as the incorporation ofsolubility enhancing agents.

Formulations for parenteral administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed, sustained, pulsed, controlled, targeted and programmed release.Thus, compounds of the invention may be formulated as a solid, semisolid, or thixotropic liquid for administration as an implanted depotproviding modified release of the active compound. Examples of suchformulations include drug coated stents and PGLA microspheres.

The compounds of the invention may also be administered topically to theskin or mucosa, that is, dermally or transdermally, such as viatransdermal patches or iontophoresis devices, intraocularadministration, or intranasal or inhalation administration. A topicalformulation may include a compound which enhances absorption orpenetration of the active ingredient through the skin or other affectedareas. When the compounds of this invention are administered by atransdermal device, administration will be accomplished using a patcheither of the reservoir and porous membrane type or of a solid matrixvariety. Typical formulations for this purpose include gels, hydrogels,lotions, solutions, creams, ointments, dusting powders, dressings,foams, films, skin patches, wafers, implants, sponges, fibers, bandagesand microemulsions. Liposomes may also be used. Typical carriers includealcohol, water, mineral oil, liquid petrolatum, white petrolatum,glycerin, polyethylene glycol and propylene glycol. Penetrationenhancers may be incorporated. Other means of topical administrationinclude delivery by electroporation, iontophoresis, phonophoresis,sonophoresis and micro needle or needle free (e.g. Powderject™,Bioject™, etc.) injection.

Formulations suitable for topical administration to the eye include, forexample, eye drops wherein the compound of this invention is dissolvedor suspended in a suitable excipient. A typical formulation suitable forocular or aural administration may be in the form of drops of amicronized suspension or solution in isotonic, pH-adjusted, sterilesaline. Other formulations suitable for ocular and aural administrationinclude ointments, biodegradable (i.e., absorbable gel sponges,collagen) and non-biodegradable (i.e., silicone) implants, wafers,lenses and particulate or vesicular systems, such as niosomes orliposomes. A polymer such as crossed linked polyacrylic acid, polyvinylalcohol, hyaluronic acid, a cellulosic polymer, for example,hydroxypropylmethylcellulose, hydroxyethylcellulose, or methylcellulose,or a heteropolysaccharide polymer, for example, gelan gum, may beincorporated together with a preservative, such as benzalkoniumchloride. Such formulations may also be delivered by iontophoresis.

Formulations for topical administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed, sustained, pulsed, controlled, targeted and programmed release.

The compounds of the invention can also be administered intranasally orby inhalation, typically in the form of a dry powder (either alone, as amixture, for example, in a dry blend with lactose, or as a mixedcomponent particle, for example, mixed with phospholipids, such asphosphatidylcholine) from a dry powder inhaler or as an aerosol sprayfrom a pressurized container, pump, spray, atomizer (preferably anatomizer using electrohydrodynamics to produce a fine mist), ornebulizer, with or without the use of a suitable propellant, such as1,1,1,2 tetrafluoroethane or 1,1,1,2,3,3,3 heptafluoropropane. Forintranasal use, the powder may include a bioadhesive agent, for example,chitosan or cyclodextrin. The pressurized container, pump, spray,atomizer, or nebulizer contains a solution or suspension of thecompound(s) of the invention comprising, for example, ethanol, aqueousethanol, or a suitable alternative agent for dispersing, solubilizing,or extending release of the active, a propellant(s) as solvent and anoptional surfactant, such as sorbitan trioleate, oleic acid, or anoligolactic acid. Prior to use in a dry powder or suspensionformulation, the drug product may be micronized to a size suitable fordelivery by inhalation (typically less than 5 microns). This may beachieved by any appropriate comminuting method, such as spiral jetmilling, fluid bed jet milling, supercritical fluid processing to formnanoparticles, high pressure homogenization, or spray drying.

Capsules (made, for example, from gelatin or HPMC), blisters andcartridges for use in an inhaler or insufflator may be formulated tocontain a powder mix of the compound of the invention, a suitable powderbase such as lactose or starch and a performance modifier such as lleucine, mannitol, or magnesium stearate. The lactose may be anhydrousor in the form of the monohydrate, preferably the latter. Other suitableexcipients include dextran, glucose, maltose, sorbitol, xylitol,fructose, sucrose and trehalose.

Suitable flavors, such as menthol and levomenthol, or sweeteners, suchas saccharin or saccharin sodium, may be added to those formulations ofthe invention intended for inhaled/intranasal administration.

Compounds of the invention may be administered rectally or vaginally,for example, in the form of a suppository, pessary, or enema. Cocoabutter is a traditional suppository base, but various alternatives maybe used as appropriate.

Formulations for rectal/vaginal administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed, sustained, pulsed, controlled, targeted and programmed release.

Compounds of the invention may also be administered directly to the eyeor ear, typically in the form of drops of a micronized suspension orsolution in isotonic, pH adjusted, sterile saline. Other formulationssuitable for ocular and aural administration include ointments,biodegradable (e.g. absorbable gel sponges, collagen) andnon-biodegradable (e.g. silicone) implants, wafers, lenses andparticulate or vesicular systems, such as niosomes or liposomes. Apolymer such as crossed linked polyacrylic acid, polyvinylalcohol,hyaluronic acid, a cellulosic polymer, for example,hydroxypropylmethylcellulose, hydroxyethylcellulose, or methylcellulose, or a heteropolysaccharide polymer, for example, gelan gum,may be incorporated together with a preservative, such as benzalkoniumchloride. Such formulations may also be delivered by iontophoresis.

Formulations for ocular/aural administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed, sustained, pulsed, controlled, targeted, or programmed release.

Other excipients and modes of administration known in the pharmaceuticalart may also be used. Pharmaceutical compositions of the invention maybe prepared by any of the well-known techniques of pharmacy, such aseffective formulation and administration procedures. The aboveconsiderations in regard to effective formulations and administrationprocedures are well known in the art and are described in standardtextbooks. Formulation of drugs is discussed in, for example, Hoover,John E., Remington's Pharmaceutical Sciences, Mack Publishing Co.,Easton, Pennsylvania, 1975; Liberman et al., Eds., Pharmaceutical DosageForms, Marcel Decker, New York, N.Y., 1980; and Kibbe et al., Eds.,Handbook of Pharmaceutical Excipients (3rd Ed.), American PharmaceuticalAssociation, Washington, 1999.

Acceptable excipients are nontoxic to subjects at the dosages andconcentrations employed, and may comprise one or more of thefollowing: 1) buffers such as phosphate, citrate, or other organicacids; 2) salts such as sodium chloride; 3) antioxidants such asascorbic acid or methionine; 4) preservatives such asoctadecyldimethylbenzyl ammonium chloride, hexamethonium chloride,benzalkonium chloride, benzethonium chloride, phenol, butyl or benzylalcohol; 5) alkyl parabens such as methyl or propyl paraben, catechol,resorcinol, cyclohexanol, 3-pentanol, or m-cresol; 6) low molecularweight (less than about 10 residues) polypeptides; 7) proteins such asserum albumin, gelatin, or immunoglobulins; 8) hydrophilic polymers suchas polyvinylpyrrolidone; 9) amino acids such as glycine, glutamine,asparagine, histidine, arginine, or lysine; 10) monosaccharides,disaccharides, or other carbohydrates including glucose, mannose, ordextrins; 11) chelating agents such as EDTA; 12) sugars such as sucrose,mannitol, trehalose or sorbitol; 13) salt-forming counter-ions such assodium, metal complexes (e.g., Zn-protein complexes), or 14) non-ionicsurfactants such as polysorbates (e.g., polysorbate 20 or polysorbate80), poloxamers or polyethylene glycol (PEG).

Liposome containing compounds of the invention may be prepared bymethods known in the art (See, for example, Chang, H. I.; Yeh, M. K.;Clinical development of liposome-based drugs: formulation,characterization, and therapeutic efficacy; Int J Nanomedicine 2012; 7;49-60). Particularly useful liposomes may be generated by the reversephase evaporation method with a lipid composition comprisingphosphatidylcholine, cholesterol and PEG-derivatizedphosphatidylethanolamine (PEG-PE). Liposomes are extruded throughfilters of defined pore size to yield liposomes with the desireddiameter.

Compounds of the invention may also be entrapped in microcapsulesprepared, for example, by coacervation techniques or by interfacialpolymerization, for example, hydroxymethylcellulose orgelatin-microcapsules and poly-(methylmethacrylate) microcapsules,respectively, in colloidal drug delivery systems (for example,liposomes, albumin microspheres, microemulsions, nano-particles andnanocapsules) or in macroemulsions. Such techniques are disclosed inRemington, The Science and Practice of Pharmacy, 20th Ed., MackPublishing (2000).

Sustained-release preparations may be used. Suitable examples ofsustained-release preparations include semi-permeable matrices of solidhydrophobic polymers containing a compound of the invention, whichmatrices are in the form of shaped articles, e.g., films, ormicrocapsules. Examples of sustained-release matrices includepolyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate),or ‘poly(vinylalcohol)), polylactides, copolymers of L-glutamic acid and7 ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradablelactic acid-glycolic acid copolymers such as those used in leuprolideacetate for depot suspension (injectable microspheres composed of lacticacid-glycolic acid copolymer and leuprolide acetate), sucrose acetateisobutyrate, and poly-D-(−)-3-hydroxybutyric acid.

The formulations to be used for intravenous administration must besterile. This is readily accomplished by, for example, filtrationthrough sterile filtration membranes. Compounds of the invention aregenerally placed into a container having a sterile access port, forexample, an intravenous solution bag or vial having a stopper pierceableby a hypodermic injection needle.

Suitable emulsions may be prepared using commercially available fatemulsions, such as a lipid emulsions comprising soybean oil, a fatemulsion for intravenous administration (e.g., comprising safflower oil,soybean oil, egg phosphatides and glycerin in water), emulsionscontaining soya bean oil and medium-chain triglycerides, and lipidemulsions of cottonseed oil. The active ingredient may be eitherdissolved in a pre-mixed emulsion composition or alternatively it may bedissolved in an oil (e.g., soybean oil, safflower oil, cottonseed oil,sesame oil, corn oil or almond oil) and an emulsion formed upon mixingwith a phospholipid (e.g., egg phospholipids, soybean phospholipids orsoybean lecithin) and water. It will be appreciated that otheringredients may be added, for example glycerol or glucose, to adjust thetonicity of the emulsion. Suitable emulsions will typically contain upto 20% oil, for example, between 5 and 20%. The fat emulsion maycomprise fat droplets between 0.1 and 1.0 μm, particularly 0.1 and 0.5μm, and have a pH in the range of 5.5 to 8.0.

For example, the emulsion compositions may be those prepared by mixing acompound of the invention with a lipid emulsions comprising soybean oilor the components thereof (soybean oil, egg phospholipids, glycerol andwater).

A drug product intermediate (DPI) is a partly processed material thatmust undergo further processing steps before it becomes bulk drugproduct. Compounds of the invention may be formulated into drug productintermediate DPI containing the active ingredient in a higher freeenergy form than the crystalline form. One reason to use a DPI is toimprove oral absorption characteristics due to low solubility, slowdissolution, improved mass transport through the mucus layer adjacent tothe epithelial cells, and in some cases, limitations due to biologicalbarriers such as metabolism and transporters. Other reasons may includeimproved solid state stability and downstream manufacturability. In oneembodiment, the drug product intermediate contains a compound of theinvention isolated and stabilized in the amorphous state (for example,amorphous solid dispersions (ASDs)). There are many techniques known inthe art to manufacture ASD's that produce material suitable forintegration into a bulk drug product, for example, spray drieddispersions (SDD's), melt extrudates (often referred to as HME's),co-precipitates, amorphous drug nanoparticles, and nano-adsorbates. Inone embodiment amorphous solid dispersions comprise a compound of theinvention and a polymer excipient. Other excipients as well asconcentrations of said excipients and the compound of the invention arewell known in the art and are described in standard textbooks. See, forexample, “Amorphous Solid Dispersions Theory and Practice” by NavnitShah et al.

In another aspect, the invention provides a compound of the invention,or a pharmaceutically acceptable salt thereof, for use as a medicament,in particular a medicament for the treatment of abnormal cell growth.

In yet another aspect, the invention provides the use of a compound ofthe invention, or a pharmaceutically acceptable salt thereof, formanufacture of a medicament for the treatment of abnormal cell growth ina subject, for example a tumor, for example a MEK-associated tumor, in asubject.

In yet another aspect, the invention provides a compound according toany of the formulae described herein, or a pharmaceutically acceptablesalt thereof, for use in the treatment of abnormal cell growth, forexample a tumor, for example a MEK-associated tumor.

Administration of the compounds of the invention may be affected by anymethod that enables delivery of the compounds to the site of action.These methods include oral routes, intraduodenal routes, parenteralinjection (including intravenous, subcutaneous, intramuscular,intravascular or infusion), topical, and rectal administration.

Dosage regimens may be adjusted to provide the optimum desired response.For example, a single bolus may be administered, several divided dosesmay be administered over time or the dose may be proportionally reducedor increased as indicated by the exigencies of the therapeuticsituation. It is especially advantageous to formulate parenteralcompositions in dosage unit form for ease of administration anduniformity of dosage. Dosage unit form, as used herein, refers tophysically discrete units suited as unitary dosages for the mammaliansubjects to be treated; each unit containing a predetermined quantity ofactive compound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical carrier. The specificationfor the dosage unit forms of the invention are dictated by and directlydependent on (a) the unique characteristics of the compound administeredand the particular therapeutic or prophylactic effect to be achieved,and (b) the limitations inherent in the art of compounding such anactive compound for the treatment of sensitivity in individuals.

Thus, the skilled artisan would appreciate, based upon the disclosureprovided herein, that the dose and dosing regimen is adjusted inaccordance with methods well known in the therapeutic arts. That is, themaximum tolerable dose can be readily established, and the effectiveamount providing a detectable therapeutic benefit to a patient may alsobe determined, as can the temporal requirements for administering eachagent to provide a detectable therapeutic benefit to the patient.Accordingly, while certain dose and administration regimens areexemplified herein, these examples in no way limit the dose andadministration regimen that may be provided to a patient in practicingthe present invention.

It is to be noted that dosage values may vary with the type and severityof the condition to be alleviated and may include single or multipledoses. It is to be further understood that for any particular subject,specific dosage regimens should be adjusted over time according to theindividual need and the professional judgment of the personadministering or supervising the administration of the compositions, andthat dosage ranges set forth herein are exemplary only and are notintended to limit the scope or practice of the claimed composition. Forexample, doses may be adjusted based on pharmacokinetic orpharmacodynamic parameters, which may include clinical effects such astoxic effects and/or laboratory values. Thus, the present inventionencompasses intrapatient dose escalation as determined by the skilledartisan. Determining appropriate dosages and regimens for administrationof the chemotherapeutic agent are well known in the relevant art andwould be understood to be encompassed by the skilled artisan onceprovided the teachings disclosed herein. In one embodiment, an effectivedosage is typically in the range of about 0.001 to about 100 mg per kgbody weight per day, and frequently about 0.01 to about 35 mg/kg/day, insingle or divided doses. For a 70 kg human, this would amount to about0.07 mg/day to about 7000 mg/day, more commonly, from about 10 mg/day toabout 1000 mg/day. Sometimes, the dosage is about 10, 20, 30, 40, 50,60, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400,425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, 700, 750, 800,900 or 1000 mg/day. Sometimes, the dosage is from about 10 mg/day toabout 1000 mg/day, from about 10 mg/day to about 750 mg/day, from about10 mg/day to about 600 mg/day, from about 10 mg/day to about 300 mg/day,from about 10 mg/day to about 150 mg/day, from about 20 mg/day to about750 mg/day, from about 20 mg/day to about to 600 mg/day, from about 20mg/day to about to 300 mg/day, from about 20 mg/day to about to 150mg/day, from about 50 mg/day to about 750 mg/day, from about 50 mg/dayto about 600 mg/day, from about 50 mg/day to about 300 mg/day, fromabout 50 mg/day to about 150 mg/day, from about 75 mg/day to about 750mg/day, from about 75 mg/day to about 600 mg/day, from about 75 mg/dayto about 300 mg/day, or from about 75 mg/day to about 150 mg/day. Insome instances, dosage levels below the lower limit of the aforesaidrange may be more than adequate, while in other cases still larger dosesmay be used without causing any harmful side effect, with such largerdoses typically divided into several smaller doses for administrationthroughout the day. In one embodiment, the subject is administered about50 mg/day.

Inasmuch as it may be desirable to administer a combination of activecompounds, for example, for the purpose of treating a particular diseaseor condition, it is within the scope of the present invention that twoor more pharmaceutical compositions, at least one of which contains acompound in accordance with the invention, may conveniently be combinedin the form of a kit suitable for co-administration of the compositions.Thus, the kit of the invention includes two or more separatepharmaceutical compositions, at least one of which contains a compoundof the invention, and means for separately retaining said compositions,such as a container, divided bottle, or divided foil packet. An exampleof such a kit is the familiar blister pack used for the packaging oftablets, capsules and the like.

The kit of the invention is particularly suitable for administeringdifferent dosage forms, for example, oral and parenteral, foradministering the separate compositions at different dosage intervals,or for titrating the separate compositions against one another. Toassist compliance, the kit typically includes directions foradministration and may be provided with a memory aid. In someembodiments, the kit includes the compound or a pharmaceuticalcomposition thereof and a diagnostic agent. In other embodiments, thekit includes the compound or a pharmaceutical composition thereof andone or more therapeutic agents, such as a BRAF inhibitor, for example, aBRAF inhibitor selected fromN-(3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-2,4-difluorophenyl)propane-1-sulfonamide;N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-3-fluoropropane-1-sulfonamide;N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4,5-difluorophenyl)propane-1-sulfonamide;N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)propane-1-sulfonamide;N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoropropane-1-sulfonamide;N-(2-chloro-4-fluoro-3-((5-methyl-3-(methyl-d3)-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-phenyl)-3-fluoropropane-1-sulfonamide;N-{2-chloro-3-[(3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy]-4-fluorophenyl}propane-1-sulfonamide;N-(3-chloro-4-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy)-5-fluoropyridin-2-yl)propane-1-sulfonamide;N-{2-chloro-3-[(3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy]-4-fluorophenyl}-3-fluoropropane-1-sulfonamide;N-(2-chloro-4-fluoro-3-((5-fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-2-azabicyclo[2.1.1]hexane-2-sulfonamide,(R)-N-(2-chloro-4-fluoro-3-((5-fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-3-fluoropyrrolidine-1-sulfonamide,andN-(2-chloro-3-((5-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoroazetidine-1-sulfonamide;or a pharmaceutically acceptable salt thereof;

The following schemes and written descriptions provide general detailsregarding the preparation of the compounds of the invention.

The compounds of the invention may be prepared by any method known inthe art for the preparation of compounds of analogous structure. Inparticular, the compounds of the invention can be prepared by theprocedures described by reference to the Schemes that follow, or by thespecific methods described in the Examples, or by similar processes toeither.

The skilled person will appreciate that the experimental conditions setforth in the schemes that follow are illustrative of suitable conditionsfor effecting the transformations shown, and that it may be necessary ordesirable to vary the precise conditions employed for the preparation ofcompounds of Formula I, and compounds that fall within Formula I, e.g.,compounds of Formula II and the like.

In addition, the skilled person will appreciate that it may be necessaryor desirable at any stage in the synthesis of compounds of the inventionto protect one or more sensitive groups, to prevent undesirable sidereactions. In particular, it may be necessary or desirable to protectamino or alcohol groups. The protecting groups (PGs) used in thepreparation of the compounds of the invention may be used inconventional manner. See, for example, those described in ‘Greene'sProtective Groups in Organic Synthesis’ by Theodora W Greene and Peter GM Wuts, third edition, (John Wiley and Sons, 1999), in particularchapters 7 (“Protection for the Amino Group”) and 2 (“Protection for theHydroxyl Group, Including 1,2- and 1,3-Diols”), incorporated herein byreference, which also describes methods for the removal of such groups.

Scheme 1 describes a general method for preparing compound 8, which is acompound of Formula I wherein R¹ is H, R² is H, R³ is C3-C6 cycloalkyl,and R⁴ is as defined for Formula I. Commercially available2,6-dichloro-4-methylnicotinic acid (compound 1) may be converted to theester analog 2 upon treatment with (trimethylsilyl)diazomethane.Compound 2 may be converted to the dimethylaminovinyl intermediatecompound 3 upon treatment with N,N-dimethylformamide dimethylacetal.Compound 3 may be converted to the aldehyde intermediate compound 4 upontreatment with a suitable acid such as hydrochloric acid in a suitablesolvent such as ether. Cyclization of compound 4 may be achieved bytreatment with compound 4 with a reagent having the formula R³NH₂,wherein R³ is C3-C6 cycloalkyl, in the presence of a reducing agent(e.g., sodium cyanoborohydride), in a suitable solvent such as methanolto provide compound 5. Compound 5 may be converted to compound 6 upontreatment with trimethylsilyl iodide in a suitable solvent, such asacetonitrile. Compound 6 may be methylated by treatment with methyliodide in the presence of a suitable base, such as an alkalinecarbonate, such as potassium carbonate, in the presence of a suitablesolvent such as THF, to provide compound 7. Compound 7 may undergoaromatic nucleophilic substitution upon treatment with a reagent havingthe formula R⁴NH₂ wherein R⁴ is as defined for Formula I, in thepresence of a strong base, such as lithium hexamethyldisilazide in asuitable solvent, such as THF, to provide compound 8.

Scheme 2 describes a general method for preparing compound 17, which isa compound of Formula I wherein R¹ is H, R² is H, R³ is hydroxy-C1-C6alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6alkoxy-, and R⁴ is as defined for Formula I. Commercially available4-bromo-2,6-dichloropyridine may be lithiated with a reagent such aslithium diisopropylamide and trapped with carbon dioxide to affordcarboxylic acid 10. Compound 10 may be converted to compound 11 byrefluxing in an aqueous base such as 4M sodium hydroxide. Compound 11may be methylated by treatment with methyl iodide in the presence of asuitable base, such as an alkaline carbonate, such as potassiumcarbonate, in the presence of a suitable solvent such as DMF, to providecompound 12. Compound 12 may be converted to vinyl ether intermediate 13via Suzuki reaction with(E)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (compound(i)) using a catalyst such asmethanesulfonato(2-dicyclohexylphosphino-2′,6′-di-i-propoxy-1,1′-biphenyl)(2-amino-1,1′-biphenyl-2-yl)palladium(II)and alkaline base (e.g., an alkaline carbonate, e.g., aqueous potassiumcarbonate) in a suitable solvent such as 1,4-dioxane. Compound 13 mayundergo oximine formation upon treatment with a reagent of formulaR^(3a)—NH₂HCl, wherein R^(3a) is P¹O—C1-C6 alkoxy-, C1-C6 alkoxy,fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy- and P¹ is analcohol protecting group such as tert-butyl, benzyl ortert-butyldimethylsilyl, using triethylamine and HCl and heating in asuitable solvent such as 1,4-dioxane to provide compound 14. Compound 14may be reduced to alkoxyamine intermediate 15 using an appropriatereducing agent such as sodium cyanoborohydride in a suitable solventsuch as isopropanol. Compound 15 may undergo aromatic nucleophilicsubstitution upon treatment with a reagent having the formula R⁴NH₂wherein R⁴ is as defined for Formula I in the presence of a strong base,such as lithium hexamethyldisilazide in a suitable solvent, such as THFand concomitantly cyclized, followed by optional deprotection ifcompound 16 contains a P¹ protecting group (using standard alcoholdeprotection conditions known to persons skilled in the art, such asphosphoric acid, trifluoroacetic acid or tetrabutylammonium fluoride) toprovide compound 17 wherein R³ is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy,fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy-.

Scheme 3 describes a general method for preparing compound 20, which isa compound of Formula I wherein R¹ is H, R² is CH₃—, R³ is hydroxy-C1-C6alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6alkoxy-, and R⁴ is as defined for Formula I. Compound 15, prepared asdescribed in Scheme 2, wherein R^(3a) is P¹O—C1-C6 alkoxy-, C1-C6alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy- and P¹ isan alcohol protecting group such as tert-butyl, benzyl ortert-butyldimethylsilyl, and wherein R⁴ is as defined for Formula I, maybe cyclized upon treatment with a reagent having the formula R⁴NH₂wherein R⁴ is as defined for Formula I in the presence of a strong base,such as lithium hexamethyldisilazide, in a suitable solvent, such asTHF, to provide compound 16. Compound 16 may be iodinated usingn-iodosuccinimide and p-toluenesulfonic acid in a suitable solvent suchas 1:1 MeOH:THF, to provide compound 18. Compound 18 may undergo Negishicoupling with methylzinc(II)chloride using a catalyst, e.g., a palladiumcatalyst such as bis(tri-t-butylphosphine)palladium (0) in a suitablesolvent such as THF, followed by optional deprotection if compound 19contains a protecting group (using standard alcohol deprotectionconditions known to persons skilled in the art, such as phosphoric acid,trifluoroacetic acid or tetrabutylammonium fluoride) to provide compound20 wherein R³ is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy.

Scheme 4 describes a general method for preparing compound 24, which isa compound of Formula I wherein R¹ is H, R² is H, R³ is H, and R⁴ is asdefined for Formula I. Compound 13, prepared as described in Scheme 2,may undergo aromatic nucleophilic substitution upon treatment with areagent having the formula R⁴NH₂ wherein R⁴ is as defined for Formula Iin the presence of a strong base, such as lithium hexamethyldisilazide,in a suitable solvent, such as THF, to provide compound 21. Compound 21may be hydrolyzed to aldehyde intermediate 22 under acidic conditionsusing an acid such as trifluoroacetic acid in a suitable solvent such asdichloromethane. Compound 22 may be cyclized upon treatment with areagent having the formula P²—NH₂ where P² is an amine protecting groupsuch as benzyl, p-methoxybenzyl or 2,4-dimethoxybenzyl, using a reducingagent such as sodium triacetoxyborohydride in a suitable solvent such asdichloroethane to provide compound 23. Deprotection of compound 23 maybe achieved using standard deprotection conditions known to personsskilled in the art, such as heating with trifluoroacetic acid orhydrochloric acid to provide compound 24.

Scheme 5 describes a general method for preparing compound 27, which isa compound of Formula I wherein R¹ is H, R² is H, R³ is hydroxyC1-C6alkyl, and R⁴ is as defined for Formula I. Compound 13, prepared asdescribed in Scheme 2, may be hydrolyzed to aldehyde intermediate 25using a suitable acid such as trifluoroacetic acid. Compound 25 may bereacted with a reagent of formula P¹O—(C1-C6 alkyl)-ONH₂HCl, where P¹ isan alcohol protecting group such as tert-butyldimethylsilyl, using asuitable reducing agent such as sodium triacetoxyborohydride and aceticacid in a suitable solvent such as dichloroethane at 60° C. to affordcyclized, deprotected compound 26. Compound 26 may undergo aromaticnucleophilic substitution upon treatment with a reagent having theformula R⁴NH₂, wherein R⁴ is as defined for Formula I, in the presenceof a strong base, such as lithium hexamethyldisilazide in a suitablesolvent, such as THF, to provide compound 27.

Scheme 6 describes an alternative general method for preparing compound17, which is a compound of Formula I wherein R¹ is H, R² is H, R³ ishydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6cycloalkyl)C1-C6 alkoxy, and R⁴ is as defined for Formula I. Compound28, prepared according to a method similar to that described forcompound 13, may undergo aromatic nucleophilic substitution upontreatment with a reagent having the formula R⁴NH₂ wherein R⁴ is asdefined for Formula I, in the presence of a strong base, such as lithiumhexamethyldisilazide, in a suitable solvent, such as THF, to providecompound 29. Compound 29 may undergo oximine formation upon treatmentwith a reagent of formula R^(3a)NH₂ HCl, wherein R^(3a) is P¹O—C1-C6alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6alkoxy and P¹ is an alcohol protecting group such as tert-butyl, benzylor tert-butyldimethylsilyl, using triethylamine and HCl and heating in asuitable solvent such as 1,4-dioxane to provide compound 30. Compound 30may be cyclized with an appropriate reducing agent such as sodiumcyanoborohydride and acetic acid in a suitable solvent such asisopropanol, followed by optional deprotection if compound 30 contains aP¹ protecting group (using standard alcohol deprotection conditionsknown to persons skilled in the art, such as phosphoric acid,trifluoroacetic acid or tetrabutylammonium fluoride) to provide compound17 wherein R³ is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy.

Scheme 7 describes an alternative general method for preparing compound17, which is a compound of Formula I wherein R¹ is H, R² is H, R³ ishydroxyC1-C6 alkoxy, and R⁴ is as defined for Formula I. Compound 28 mayundergo oximine formation upon treatment with a reagent of formulaTBSO-(C1-C6 alkyl)-ONH₂ HCl, using triethylamine and HCl and heating ina suitable solvent such as 1,4-dioxane to provide compound 32. Compound32 may be protected with an appropriate alcohol protecting group P¹ suchas tert-butyldimethylsilyl, using tert-butylsimethylsilyl chloride and asuitable base such as imidazole, in a suitable solvent such as DMF, toprovide compound 14. Compound 14 may be reduced to alkoxyamineintermediate 15 using an appropriate reducing agent such as sodiumcyanoborohydride in a suitable solvent such as isopropanol. Compound 15may undergo aromatic nucleophilic substitution upon treatment with areagent having the formula R⁴NH₂ wherein R⁴ is as defined for Formula Iin the presence of a strong base, such as lithium hexamethyldisilazidein a suitable solvent, such as THF, to provide compound 16. Compound 16may be deprotected using standard alcohol deprotection conditions knownto persons skilled in the art, such as phosphoric acid, trifluoroaceticacid or tetrabutylammonium fluoride to provide compound 17.

Scheme 8 describes an alternative general method for preparing compound20, which is a compound of Formula I wherein R¹ is H, R² is CH₃—, R³ ishydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6cycloalkyl)C1-C6 alkoxy, and R⁴ is as defined for Formula I. Compound 28may undergo aromatic nucleophilic substitution upon treatment with areagent having the formula R⁴NH₂ wherein R⁴ is as defined for Formula I,in the presence of a strong base, such as lithium hexamethyldisilazide,in a suitable solvent, such as THF, to provide compound 29. Compound 29may be hydrolyzed to aldehyde intermediate 22 using a suitable acid suchas trifluoroacetic acid. Compound 22 may be reacted with a reagent offormula R^(3I)NH₂ HCl, wherein R^(3a) is P¹O—C1-C6 alkoxy-, C1-C6alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy and P¹ isan alcohol protecting group such as tert-butyl, benzyl ortert-butyldimethylsilyl, using a suitable reducing agent such as sodiumtriacetoxyborohydride and acetic acid in a solvent such asdichloroethane at 60° C. to afford cyclized compound 16. Compound 16 maybe iodinated using n-iodosuccinimide and p-toluenesulfonic acid in asuitable solvent such as 1:1 MeOH:THF, to provide compound 18. Compound18 may undergo Negishi coupling with methylzinc(II)chloride using acatalyst such as bis(tri-t-butylphosphine)palladium (0) in a suitablesolvent such as THF, followed by optional deprotection if compound 18contains a P¹ protecting group (using standard alcohol deprotectionconditions known to persons skilled in the art, such as phosphoric acid,trifluoroacetic acid or tetrabutylammonium fluoride) to provide compound20 wherein R³ is hydroxy-C1-C6 alkoxy-C1-C6 alkoxy, fluoroC1-C6 alkoxy,or (C3-C6 cycloalkyl)C1-C6 alkoxy.

Scheme 9 describes a general process for the synthesis of compound 35,which is a compound of Formula I wherein R¹ is H, R² is halogen, R³ ishydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6cycloalkyl)C1-C6 alkoxy, and R⁴ is as defined for Formula I. Compound 12may undergo aromatic nucleophilic substitution upon treatment with areagent having the formula R⁴NH₂ wherein R⁴ is as defined for Formula I,in the presence of a strong base, such as lithium hexamethyldisilazide,in a suitable solvent, such as THF, to provide compound 33. Compound 33may be converted to vinyl ether intermediate 21 via Suzuki reaction with(E)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane using acatalyst, e.g., a palladium catalyst such asmethanesulfonato(2-dicyclohexylphosphino-2′,6′-di-i-propoxy-1,1′-biphenyl)(2-amino-1,1-biphenyl-2-yl)palladium(II)and alkaline base, e.g., an alkaline carbonate base such as aqueouspotassium carbonate, in a suitable solvent such as 1,4-dioxane. Compound21 may undergo oximine formation upon treatment with a reagent offormula R^(3a)NH₂ HCl, wherein R^(3a) is P¹O—C1-C6 alkoxy-, C1-C6alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy and P¹ isan alcohol protecting group such as tert-butyl, benzyl ortert-butyldimethylsilyl, using triethylamine and HCl and heating in asuitable solvent such as 1,4-dioxane to provide compound 30. Compound 30may be treated with an appropriate reducing agent such as sodiumcyanoborohydride and acetic acid in a suitable solvent such asisopropanol, to generate cyclized compound 31. Compound 31 may behalogenated using conditions such as treatment with N-iodosuccinimideand p-toluenesulfonic acid in 1:1 THF/MeOH, or N-bromosuccinimide in asuitable solvent such as DMF, or N-chlorosuccinimide in a suitablesolvent such as DMF, or Selectfluor in a suitable solvent such asacetonitrile, followed by optional deprotection if compound 31 has a P¹protecting group (such as phosphoric acid, trifluoroacetic acid ortetrabutylammonium fluoride) to provide compound 35 wherein R² is iodo,bromo, chloro or fluoro, respectively, and R³ is hydroxy-C1-C6 alkoxy-,C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy

Scheme 10 describes an alternative general method for preparing compound17, which is a compound of Formula I wherein R¹ is H, R² is H, R³ ishydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6cycloalkyl)C1-C6 alkoxy, and R⁴ is as defined for Formula I. Compound 12may undergo aromatic nucleophilic substitution upon treatment with areagent having the formula R⁴NH₂ wherein R⁴ is as defined for Formula I,in the presence of a suitable base, such as potassium tert-butoxide in asuitable solvent, such as THF, to provide compound 36. Compound 36 maybe converted to vinyl ether intermediate 29 via Suzuki reaction with(Z)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane using acatalyst, e.g., a palladium catalyst such asmethanesulfonato(2-dicyclohexylphosphino-2′,6′-di-i-propoxy-1,1′-biphenyl)(2-amino-1,1′-biphenyl-2-yl)palladium(II)and suitable base such as an alkaline base, e.g., an alkaline carbonatebase, e.g., aqueous potassium carbonate in a solvent such as2-methyltetrahydrofuran. Compound 29 may be reacted with a reagent offormula R^(3a)NH₂ HCl, wherein R^(3a) is P¹O—C1-C6 alkoxy-, C1-C6alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy and P¹ isan alcohol protecting group such as tert-butyl, benzyl ortert-butyldimethylsilyl, using triethylamine and hydrochloric acid in asuitable solvent such as 1,4-dioxane, then treated with a suitablereducing agent such as pyridine borane and hydrochloric acid and heatedat 60° C., followed by optional deprotection if compound 29 has a P¹protecting group (such as phosphoric acid, trifluoroacetic acid ortetrabutylammonium fluoride) to provide compound 17 wherein R³ ishydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6cycloalkyl)C1-C6 alkoxy.

Scheme 11 describes a process for preparing compound 43, which is acompound of Formula I wherein R¹ is phenyl, R² is hydrogen, R³ ishydrogen, and R⁴ is as described for Formula I. Commercially available2,6-dichloro-4-iodopyridine may be lithiated with a reagent such aslithium diisopropylamide and trapped with carbon dioxide to affordcompound 37. Compound 37 may be converted to compound 38 by refluxing inan aqueous base such as 4M sodium hydroxide. Compound 38 may bemethylated by treatment with methyl iodide in the presence of a suitablebase, such as an alkaline carbonate, such as potassium carbonate, in thepresence of a suitable solvent such as DMF, to provide compound 39.Compound 39 may be converted to vinyl ether intermediate 40 via Suzukireaction with 4,4,5,5-tetramethyl-2-(1-phenylvinyl)-1,3,2-dioxaborolaneusing a suitable catalyst, for example a palladium catalyst (for examplePd(dppf)Cl₂) and base such as such as an alkaline carbonate (for exampleaqueous potassium carbonate) in a suitable solvent such as 1,4-dioxane.Compound 40 may undergo aromatic nucleophilic substitution upontreatment with a reagent having the formula R⁴NH₂, wherein R⁴ is asdefined for Formula I, in the presence of a strong base, such as lithiumhexamethyldisilazide, in a suitable solvent such as THF, to providecompound 41. Compound 41 may be cyclized with(2,4-dimethoxyphenyl)methanamine in the presence of a Lewis acid such astrimethylaluminum by heating in a suitable solvent such as toluene toafford compound 42. Compound 42 may be deprotected by heating with asuitable acid such as TFA to afford compound 43.

Scheme 12 describes a method of preparing compound 51, which is acompound of Formula I wherein R¹ is methyl, R² is hydrogen, R³ ishydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6cycloalkyl)C1-C6 alkoxy, and R⁴ is as defined for Formula I. Compound 44may be treated with NH₃, Fe(NO₃)₃·9H₂O and NaNH₂ at low temperature,followed by treatment with methyl iodide to provide compound 45.Compound 45 can be heated with benzo[d][1,3,2]dioxaborole in a suitablesolvent such as toluene with a catalyst such as NiCl₂(dppe) to providecompound 46. Compound 46 may be converted to compound 47 by reactionwith compound 39 using Suzuki reaction conditions, for example in thepresence of a catalyst, for example a palladium catalyst such asPd(dppf)Cl₂, and base such as K₃PO₄, K₂CO₃, KOtBu, Cs₂CO₃, NaOH, ortriethylamine, in a suitable solvent, for example 1 solvent mixture suchas toluene/THF. Compound 47 may undergo oximine formation upon treatmentwith a reagent of formula R^(3a)NH₂ HCl, wherein R³¹ is P¹O—C1-C6alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6alkoxy and P¹ is an alcohol protecting group such as tert-butyl, benzylor tert-butyldimethylsilyl, using triethylamine and HCl and heating in asuitable solvent such as 1,4-dioxane to provide compound 48. Compound 48may be treated with an appropriate reducing agent such as sodiumcyanoborohydride and acetic acid in a suitable solvent such asisopropanol, to generate cyclized product 49. Compound 49 may undergoaromatic nucleophilic substitution upon treatment with a reagent havingthe formula R⁴NH₂, wherein R⁴ is as defined for Formula I, in thepresence of a strong base, such as lithium hexamethyldisilazide in asuitable solvent, such as THF, followed by optional deprotection ifcompound 49 contains a P¹ protecting group (using standard alcoholdeprotection conditions known to persons skilled in the art, such asphosphoric acid, trifluoroacetic acid or tetrabutylammonium fluoride) toprovide compound 51 wherein R³ is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy,fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy.

The term “amine protecting group” as used herein refers to a derivativeof the groups commonly employed to block or protect an amino group whilereactions are carried out on other functional groups on the compound.Examples of suitable protecting groups for use in any of the processesdescribed herein include carbamates, amides, alkyl and aryl groups,imines, as well as many N-heteroatom derivatives which can be removed toregenerate the desired amine group. Non-limiting examples of amineprotecting groups are t-butyloxycarbonyl (“Boc”),2-trimethylsilylethoxymethyl (SEM), and p-methoxybenzyl (PMB). Furtherexamples of these groups, and other protecting groups, are found in T.W. Greene, et al., Greene's Protective Groups in Organic Synthesis. NewYork: Wiley Interscience, 2006.

The term “alcohol protecting group” as used herein refers to aderivative of the groups commonly employed to block a hydroxy groupwhile reactions are carried out on other functional groups on thecompound. Examples of suitable protecting groups for use in any of theprocesses described herein include benzyl, trityl, silyl ethers, and thelike.

Intermediate compounds 7, 15, 18, 22, 26, 29, 30, and 31, as illustratedin the above Schemes, are also novel intermediates useful for thepreparation of compounds of Formula I and provide further embodiments ofthe invention.

In one embodiment, provided herein is a process for preparing compoundsof Formula I, comprising:

-   -   (a) for a compound of Formula I wherein R¹ is H, R² is H, R³ is        C3-C6 cycloalkyl, and R⁴ is as defined for Formula I, reacting a        compound of formula 7

with a reagent having the formula R⁴NH₂ wherein R⁴ is as defined forFormula I, in the presence of a strong base; or

-   -   (b) for a compound of Formula I wherein R¹ is H, R² is H, R³ is        hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or        (C3-C6 cycloalkyl)C1-C6 alkoxy-, and R⁴ is as defined for        Formula I, cyclizing a compound of formula 15

wherein R^(3a) is P¹O—C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy,or (C3-C6 cycloalkyl)C1-C6 alkoxy- and P¹ is an alcohol protectinggroup, in the presence of a reagent having the formula R⁴NH₂ wherein R⁴is as defined for Formula I, in the presence of a strong base,optionally followed by removal of the alcohol protecting group ifpresent; or

-   -   (c) for a compound of Formula I wherein R¹ is H, R² is CH₃—, R³        is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or        (C3-C6 cycloalkyl)C1-C6 alkoxy-, and R⁴ is as defined for        Formula I, treating a compound of formula 18

wherein R^(3a) is P¹O—C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy,or (C3-C6 cycloalkyl)C1-C6 alkoxy, P¹ is an alcohol protecting group,and R⁴ is as defined for Formula I, with methylzinc(II) chloride in thepresence of a palladium catalyst, followed by removal of the alcoholprotecting group if present; or

-   -   (d) for a compound of Formula I wherein R¹ is H, R² is H, R³ is        H, and R⁴ is as defined for Formula I, cyclizing a compound of        formula 22

wherein R⁴ is as defined for Formula I, in the presence of a reagenthaving the formula P²—NH₂ where P² is an amine protecting group,followed by removing the amine protecting group; or

-   -   (e) for a compound of Formula I wherein R¹ is H, R² is H, R³ is        hydroxyC1-C6 alkyl, and R⁴ is as defined for Formula I, reacting        a compound of the formula 26

-   -   in the presence of a reagent having the formula R⁴NH₂ in the        presence of a strong base, wherein R⁴ is as defined for Formula        I; or    -   (f) for a compound of Formula I wherein R¹ is H, R² is H, R³ is        hydroxyC1-C6 alkyl, and R⁴ is as defined for Formula I,        cyclizing a compound having formula 30

wherein P¹ is an alcohol protecting group and R⁴ is as defined forFormula I, in the presence of a reducing agent, followed by removal ofthe alcohol protecting group; or

-   -   (g) for a compound of Formula I wherein R¹ is H, R² is halogen,        R³ is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy,        or (C3-C6 cycloalkyl)C1-C6 alkoxy-, and R⁴ is as defined for        Formula I, halogenating a compound of formula 31

wherein R⁴ is as defined for Formula I, R^(3a) is P¹O—C1-C6 alkoxy-,C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy- andP¹ is an alcohol protecting group, followed by removal of the alcoholprotecting group if present; or

-   -   (h) for a compound of Formula I wherein R¹ is H, R² is H, R³ is        hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or        (C3-C6 cycloalkyl)C1-C6 alkoxy-, and R⁴ is as defined for        Formula I, reacting a compound of formula 29

wherein R⁴ is as defined for Formula I, with a reagent of formulaP¹O—(C1-C6 alkyl)-ONH₂HCl, where P¹ is an alcohol protecting group, inthe presence of triethylamine and hydrochloric acid, followed by removalof the alcohol protecting group; or

-   -   (i) for a compound of Formula I wherein R¹ is phenyl, R² is        hydrogen, R³ is hydrogen, and R⁴ is as described for Formula I,        cyclizing a compound having the formula 41

wherein R⁴ is defined as for Formula I, with(2,4-dimethoxyphenyl)methanamine in the presence of a Lewis acid at anelevated temperature to provide a compound having the formula 42:

followed by treatment of compound 42 with acid; or

-   -   (j) for a compound of Formula I wherein R¹ is methyl, R² is        hydrogen, R³ is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6        alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy and R⁴ is as defined        for Formula I, reacting a compound having the formula 49

wherein R^(3a) is P¹O—C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy,or (C3-C6 cycloalkyl)C1-C6 alkoxy- and P¹ is an alcohol protectinggroup, with a reagent having the formula R⁴NH₂, wherein R⁴ is as definedfor Formula I, in the presence of a strong base, followed by removal ofthe P¹ protecting group if present; and

-   -   optionally converting the compound of Formula I to a        pharmaceutically acceptable salt.

Synthetic intermediates 3, 4, 5, 6, 7, 13, 14, 15, 16, 18, 21, 22, 23,25, 26, 28, 29, 30, 32, 33, 36, 37, 38, 39, 40, 41, 42, 47, 48, and 49are also believed to be novel and are further embodiments of thisinvention.

The present invention may be further understood by reference to thefollowing detailed description of the embodiments of the invention andthe Examples included herein. It is to be understood that this inventionis not limited to specific synthetic methods of making that may ofcourse vary. It is to be also understood that the terminology usedherein is for the purpose of describing specific embodiments only and isnot intended to be limiting.

-   -   E1. A compound of Formula I:

-   -   or a pharmaceutically acceptable salt thereof, wherein:        -   R¹ is H, Br, C1-C6 alkyl or phenyl;        -   R² is H, halogen or CH₃—;        -   R³ is H, hydroxyC1-C6 alkyl-, hydroxyC1-C6 alkoxy-, C1-C6            alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, or (C3-C6            cycloalkyl)C1-C6 alkoxy-; and        -   R⁴ is phenyl substituted with 1, 2 or 3 substituents            independently selected from halogen, C1-C6 alkyl, C1-C6            alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6            alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, and C1-C6            alkyl-C(═O)—.    -   E2. A compound according to embodiment E1, or a pharmaceutically        acceptable salt thereof, wherein R¹ is H.    -   E3. A compound according to embodiment E1 or E2, or a        pharmaceutically acceptable salt thereof, wherein R² is H.    -   E4. A compound according to embodiment E1 or E2, or a        pharmaceutically acceptable salt thereof, wherein R² is CH₃—.    -   E5. A compound according to any one of embodiments E1 to E4 or a        pharmaceutically acceptable salt thereof, wherein R³ is        hydroxyC1-C6 alkyl-.    -   E6. A compound according to any one of embodiments E1 to E5, or        a pharmaceutically acceptable salt thereof, wherein R⁴ is phenyl        substituted with 1 or 2 substituents independently selected from        halogen, C1-C6 alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio,        fluoroC1-C6 alkyl, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6        cycloalkyl, and C1-C6 alkyl-C(═O)—.    -   E7. A compound according to any one of embodiments E1 to E6 or a        pharmaceutically acceptable salt thereof, wherein R⁴ is phenyl        substituted with 1 or 2 substituents independently selected from        halogen and C1-C6 alkylthio.    -   E8. A compound of Formula II:

-   -   or a pharmaceutically acceptable salt thereof, wherein        -   R¹ is H, Br, C1-C6 alkyl or phenyl;        -   R² is H, halogen or CH₃—;        -   R³ is H, hydroxyC1-C6 alkyl-, hydroxyC1-C6 alkoxy-, C1-C6            alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, or (C3-C6            cycloalkyl)C1-C6 alkoxy-; and        -   R^(a) and R^(b) are independently selected from halogen,            C1-C6 alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio,            fluoroC1-C6 alkyl, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6            cycloalkyl, and C1-C6 alkyl-C(═O)—.    -   E9. A compound according to embodiment E8, or a pharmaceutically        acceptable salt thereof, wherein R¹ is H.    -   E10. A compound according to embodiment E8 or E9, or a        pharmaceutically acceptable salt thereof, wherein R² is H.    -   E11. A compound according to embodiment E8 or E9, or a        pharmaceutically acceptable salt thereof, wherein R² is CH₃—.    -   E12. A compound according to any one of embodiments E8 to E11,        or a pharmaceutically acceptable salt thereof, wherein R³ is        hydroxyC1-C6 alkyl-.    -   E13. A compound according to any one of embodiments E8 to E11,        or a pharmaceutically acceptable salt thereof, wherein R³ is H.    -   E14 A compound according to any one of embodiment E8 to E13, or        a pharmaceutically acceptable salt thereof, wherein R^(a) is        halogen.    -   E15. A compound according to any one of embodiment E8 to E14, or        a pharmaceutically acceptable salt thereof, wherein R^(b) is        halogen, C1-C6 alkyl, C1-C6 alkylthio, or fluoroC1-C6 alkoxy.    -   E16. A compound according to embodiment E15, or a        pharmaceutically acceptable salt thereof, wherein R^(b) is C1-C6        alkylthio.    -   E17. A compound according to embodiment E1, selected from:

-   8-((4-bromo-2-fluorophenyl)amino)-2-cyclopropyl-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-bromo-2-fluorophenyl)amino)-2-(cyclopropylmethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-bromo-2-fluorophenyl)amino)-2-ethoxy-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   2-cyclopropyl-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   2-cyclopropyl-8-((2-fluoro-4-iodophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-chloro-4-iodophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-bromo-2-chlorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-bromo-2,3-difluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-bromo-3-chloro-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluoro-4-(trifluoromethyl)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-ethyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-cyclopropyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluoro-4-methoxyphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluoro-4-((trifluoromethyl)thio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluoro-4-isopropylphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-chloro-4-cyclopropylphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-acetyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-chloro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-chloro-4-ethylphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluoro-4-(trifluoromethoxy)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-((difluoromethyl)thio)-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-bromo-2-fluorophenyl)amino)-2-isopropoxy-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluoro-4-iodophenyl)amino)-2-isopropoxy-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   2-ethoxy-8-((2-fluoro-4-iodophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluoro-4-iodophenyl)amino)-2-methoxy-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   2-(tert-butoxy)-8-((2-fluoro-4-iodophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   (S)-8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxypropoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   (R)-8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxypropoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   (S)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxypropoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   (R)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxypropoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   (R)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxypropoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-bromo-2-fluorophenyl)amino)-5-chloro-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   5-chloro-8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-bromo-2-fluorophenyl)amino)-5-fluoro-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   5-bromo-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   4-bromo-8-((2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-ethyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-cyclopropyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-bromo-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-bromo-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluoro-4-iodophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-iodo-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-ethyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-cyclopropyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluoro-4-propylphenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluoro-4-(methylthio)phenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-ethyl-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-cyclopropyl-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-bromo-2-chlorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   2-(2,2-difluoroethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-bromo-2-fluorophenyl)amino)-7-methyl-4-phenyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-4-phenyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-bromo-2-fluorophenyl)amino)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluoro-4-(methylthio)phenyl)amino)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;

-   8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione;    -   or a pharmaceutically acceptable salt thereof.    -   E18. A compound which is        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione        having the structure:

or a pharmaceutically acceptable salt thereof.

-   -   E19. A compound which is        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione        having the structure:

-   -   E20. A crystal comprising        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.    -   E21. Crystalline anhydrous        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,        Form 1.    -   E22. Crystalline anhydrous        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,        Form 1 according to embodiment E21, having a PXRD pattern        comprising characterizing peaks at 5.0, 8.7, 9.3, 10.8, 14.5,        15.3, 18.8 and 20.5 degrees 2-theta (±0.2 degrees 2-theta).    -   E23. Crystalline anhydrous        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,        Form 1 according to embodiment E21, having a PXRD pattern        comprising peaks at 2-theta values essentially the same as shown        in FIG. 1 .    -   E24. Crystalline anhydrous        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,        Form 2.    -   E25. Crystalline anhydrous        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,        Form 2 according to embodiment E24, having a PXRD pattern        comprising characterizing peaks at 7.1, 9.4, 12.4, 12.8, 14.3,        15.6, 16.4, 17.4, 18.5, 18.9, 19.5, 19.9, 21.1, 21.4, 23.2,        23.7, 24.8, 25.6, 27.6, 30.3, 33.2, 33.5, and 37.5 degrees        2-theta (±0.2 degrees 2-theta).    -   E26. Crystalline anhydrous        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,        Form 2 according to embodiment E24, having a PXRD pattern        comprising peaks at 2-theta values essentially the same as shown        in FIG. 2 .    -   E27. Crystalline anhydrous        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,        Form 2 according Embodiment E25 or E26, wherein the PXRD pattern        is obtained by PXRD analysis conducted at 25° C. and at a        relative humidity below 10%.    -   E28. Crystalline monohydrate        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,        Form 3.    -   E29. Crystalline monohydrate        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,        Form 3 according to embodiment E28, having PXRD peaks at 13.7,        18.0 and 18.3 degrees 2-theta (±0.2 degrees 2-theta).    -   E30. Crystalline monohydrate        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,        Form 3 according to embodiment E28, having PXRD peaks at 6.9,        9.1, 13.7, 18.0 and 18.3 degrees 2-theta (±0.2 degrees 2-theta).    -   E31. Crystalline monohydrate        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,        Form 3 according to embodiment E28, having PXRD peaks, in terms        of 2-theta, at 6.9, 9.1, 11.8, 12.0, 13.7, 14.0, 15.2, 15.8,        18.0, 18.3, 19.0, 19.3, 20.2, 20.9, 21.6, 22.6, 23.6, 24.0,        24.9. 25.2, 25.8, 27.5, 28.1, 28.4, 29.8, 30.9, 31.7, 32.3 and        36.5 degrees 2-theta (±0.2 degrees 2-theta.    -   E32. Crystalline monohydrate        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,        Form 3 according to embodiment E28, having a PXRD pattern        comprising peaks at 2-theta values essentially the same as shown        in FIG. 3 .    -   E33. Crystalline monohydrate        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,        Form 3 according to any one of embodiments E29 to E32, wherein        the PXRD pattern is obtained by PXRD analysis conducted at        25° C. and at a relative humidity above 35%.    -   E34. Amorphous        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,        Form 4.    -   E35. Amorphous        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,        Form 4 according to embodiment E34, having a PXRD pattern        comprising peaks at 2-theta values essentially the same as shown        in FIG. 4 .    -   E36. A pharmaceutical composition, comprising a compound        according to any one of embodiments E1 to E35, or a        pharmaceutically acceptable salt thereof, and at least one        pharmaceutically acceptable excipient.    -   E37. A method of treating a MEK-associated tumor, comprising        administering to a subject in need thereof a therapeutically        effective amount of a compound according to any one of        embodiments E1 to E35, or a pharmaceutically acceptable salt        thereof.    -   E38. The method according to embodiment E37, wherein the tumor        has a BRAF V600 mutation selected from V600E, V600K, V600D,        V600R and V600S.    -   E39. The method according to embodiment E37 or E38, wherein the        tumor has a BRAF V600E mutation.    -   E40. The method according to any one of embodiments E37 to E39,        wherein the tumor is an extracranial tumor.    -   E41. The method according to embodiment E40, wherein the        extracranial tumor is selected from melanoma, colorectal cancer,        thyroid cancer, non-small cell lung cancer, ovarian cancer, and        neuroblastoma.    -   E42. The method according to any one of embodiments E37 to E39,        wherein the tumor is a CNS tumor.    -   E43. The method according to embodiment E42, wherein the CNS        tumor is an intracranial tumor.    -   E44. The method according to embodiment E43, wherein the        intracranial tumor is a brain cancer.    -   E45. The method according to embodiment E44, wherein the brain        cancer is a metastatic brain cancer.    -   E46. The method according to embodiment E45, wherein the        metastatic brain cancer is selected from metastatic melanoma,        metastatic colorectal cancer, metastatic non-small cell lung        cancer, metastatic thyroid cancer, and metastatic ovarian        cancer.    -   E47. The method according to embodiment E42, wherein the CNS        tumor is intracranial LMD or extracranial LMD.    -   E48. The method according to embodiment E47, wherein the LMD is        selected from metastatic melanoma, metastatic colorectal cancer,        and metastatic non-small cell lung cancer.    -   E49. The method according to embodiment E43, wherein the        intracranial tumor is a primary tumor.    -   E50. The method of embodiment E49, wherein the primary brain        tumor is a malignant tumor.    -   E51. The method according to embodiment E50, wherein the primary        brain tumor is a Grade 2 glioma, a Grade 3 glioma or a Grade 4        glioma.    -   E52. The method according to embodiment E49, wherein the primary        brain tumor is a benign tumor.    -   E53. The method according to embodiment E37, wherein the tumor        has a BRAF fusion.    -   E54. The method according to embodiment E53, wherein the tumor        has a BRAF fusion selected from KIAA11549-BRAF, MKRN1-BRAF,        TRIM24-BRAF, AGAP3-BRAF, ZC3HAV1-BRAF, AKAP9-BRAF, CCDC6-BRAF,        AGK-BRAF, EPS15-BRAF, NUP214-BRAF, ARMC10-BRAF, BTF3L4-BRAF,        GHR-BRAF, ZC3HAV1-BRAF, ZNF767-BRAF, CCDC91-BRAF, DYNC112-BRAF,        ZKSCAN1-BRAF, GTF21-BRAF, MZT1-BRAF, RAD18-BRAF, CUX1-BRAF,        SLC12A7-BRAF, MYRIP-BRAF, SND1-BRAF, NUB1-BRAF, KLHL7-BRAF,        TANK-BRAF, RBMS3-BRAF, STRN3-BRAF, STK35-BRAF, ETFA-BRAF,        SVOPL-BRAF, and JHDM1D-BRAF.    -   E55. The method according to embodiment E54, wherein the tumor        is breast carcinoma (e.g., breast invasive ductal carcinoma),        colorectal carcinoma (e.g., colon adenocarcinoma), esophageal        carcinoma (e.g., esophagus adenocarcinoma), glioma (e.g., brain        desmoplastic infantile ganglioglioma, brain pilocytic        astrocytoma, brain pleomorphic xanthoastrocytoma, spinal cord        low-grade glioma (NOS), anaplastic oligodendroglioma, anaplastic        ganglioglioma), head and neck carcinoma (e.g., head and neck        neuroendocrine carcinoma), lung carcinoma (e.g., lung        adenocarcinoma, lung non-small cell lung cancer (NOS)), melanoma        (e.g., cutaneous melanoma Spitzoid, mucosal melanoma        non-Spitzoid, cutaneous melanoma Spitzoid, unknown primary        melanoma, cutaneous melanoma non-Spitzoid), pancreatic carcinoma        (e.g., adenocarcinoma, pancreas acinar cell carcinoma),        prostatic carcinoma (e.g., prostate acinar adenocarcinoma),        sarcoma (malignant solid fibrous tumor), thyroid carcinoma        (thyroid papillary carcinoma), unknown primary carcinoma (e.g.,        unknown primary, adenocarcinoma), pleura mesothelioma, rectum        adenocarcinoma, uterus endometrial carcinoma (e.g., uterus        endometrial adenocarcinoma (NOS)), or ovary serous carcinoma.    -   E56. The method according to embodiment E37, wherein the tumor        is a BRAF wild-type tumor.    -   E57. The method according to any one of embodiments E37 to E56,        wherein the method further comprises administering one or more        additional anticancer therapies.    -   E58. The method according to embodiment E57, wherein the one or        more additional anticancer therapies is independently selected        from surgery, radiotherapy and anticancer agents.    -   E59. The method according to embodiment E58, wherein the        additional anticancer therapy is selected from one or more        anticancer agents.    -   E60. The method according to embodiment E59, wherein the        anticancer agent is selected from MEK inhibitors, BRAF        inhibitors, EGFR inhibitors, inhibitors of HER2 and/or HER3,        SHP2 inhibitors, Axl inhibitors, PI3K inhibitors, SOS1        inhibitors, signal transduction pathway inhibitors, checkpoint        inhibitors, modulators of the apoptosis pathway, cytotoxic        chemotherapeutics, angiogenesis-targeted therapies, and        immune-targeted agents including immunotherapy.    -   E61. The method of embodiment E60, where in the anticancer agent        is a BRAF inhibitor.    -   E62. The method of embodiment E61, wherein the BRAF inhibitor is        encorafenib or a pharmaceutically acceptable salt thereof.    -   E63. The method of embodiment E61, wherein the BRAF inhibitor is        selected from:

-   N-(3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-2,4-difluorophenyl)propane-1-sulfonamide;

-   N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-3-fluoropropane-1-sulfonamide;

-   N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4,5-difluorophenyl)propane-1-sulfonamide;

-   N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)propane-1-sulfonamide;

-   N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoropropane-1-sulfonamide;

-   N-(2-chloro-4-fluoro-3-((5-methyl-3-(methyl-d3)-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-phenyl)-3-fluoropropane-1-sulfonamide;

-   N-{2-chloro-3-[(3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy]-4-fluorophenyl}propane-1-sulfonamide;

-   N-(3-chloro-4-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy)-5-fluoropyridin-2-yl)propane-1-sulfonamide;    and

-   N-{2-chloro-3-[(3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy]-4-fluorophenyl}-3-fluoropropane-1-sulfonamide;    -   or a pharmaceutically acceptable salt thereof.    -   E64. The method of embodiment E63, wherein the BRAF inhibitor is        N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoropropane-1-sulfonamide        or a pharmaceutically acceptable salt thereof.    -   E65. The method of embodiment E61, wherein the BRAF inhibitor is        selected from:

-   N-(2-chloro-4-fluoro-3-((5-fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-2-azabicyclo[2.1.1]hexane-2-sulfonamide,

-   (R)-N-(2-chloro-4-fluoro-3-((5-fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-3-fluoropyrrolidine-1-sulfonamide,    and

-   N-(2-chloro-3-((5-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoroazetidine-1-sulfonamide,    -   or a pharmaceutically acceptable salt thereof.    -   E66. The method of embodiment E65, wherein the BRAF inhibitor is        N-(2-chloro-3-((5-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoroazetidine-1-sulfonamide,        or a pharmaceutically acceptable salt thereof.    -   E67. The method of embodiment E60, wherein the anticancer agent        is a SHP2 inhibitor.    -   E68. The method of embodiment E67, wherein the SHP2 inhibitor is        (S)-1′-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4′-piperidin]-1-amine        or a pharmaceutically acceptable salt thereof    -   E69. The method according to any one of embodiments E37 to E68,        wherein the compound is        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione        and wherein the subject is administered 50 mg of        8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione        once a day.    -   E70. A compound according to any one of embodiments E1 to E35,        or a pharmaceutically acceptable salt thereof, for use as a        medicament.    -   E71. A compound according to one of embodiments E1 to E35, or a        pharmaceutically acceptable salt thereof, for use in the        treatment of a MEK-associated tumor.    -   E72. Use of a compound according to any one of embodiments E1 to        E35 or a pharmaceutically acceptable salt thereof, for the        manufacture of a medicament for the treatment of a        MEK-associated tumor in a subject.

In order that this invention may be better understood, the followingexamples are set forth. These examples are for purposes of illustrationonly and are not to be construed as limiting the scope of the inventionin any manner.

The compounds and intermediates described below were named using thenaming convention provided with ChemDraw, Version 20.1.1.125 (PerkinElmer Informatics). The naming convention provided with ChemDraw,Version 20.1.1.125 is well known by those skilled in the art and it isbelieved that the naming convention provided with ChemDraw, Version20.1.1.125 generally comports with the IUPAC (International Union forPure and Applied Chemistry) recommendations on Nomenclature of OrganicChemistry and the CAS Index rules. Unless noted otherwise, all reactantswere obtained commercially without further purifications or wereprepared using methods known in the literature.

EXAMPLES Biological Examples Example A Cellular Phospho-p44/42 MAPK(Erk1/2) (Thr202/Tyr204) Assay

Inhibition of ERK1/2(Thr202/Tyr204) phosphorylation was determined bythe following cellular assay, which comprises incubating cells with acompound for 1 hour and quantifying pERK signal by In-Cell Western onfixed cells and normalizing to GAPDH signal. A375 cells were obtainedfrom the ATCC and grown in DMEM supplemented with 10% fetal bovineserum, penicillin/streptomycin, Glutamax©, non-essential amino acids,and sodium pyruvate. Cells were plated in 96-well plates at 30,000cells/well and allowed to attach overnight at 37° C./5% CO₂. Cells weretreated with compounds prepared as a 10-point, 1:3 dilution series(range: 20 μM-0.05 nM; maximum concentration varied from 20 μM-1 μM),with a final DMSO concentration of 0.5%. Control wells contained either0.5% DMSO alone (no inhibition control) or 1 μM of a potent controlcompound (complete inhibition control). After a 1-hour incubation, cellswere fixed in 3.7% formaldehyde in dPBS (Dulbecco's phosphate-bufferedsaline) at room temperature for 20 minutes. Cells were then washed withdPBS and permeabilized in 100% MeOH at room temperature for 10 minutes.Following permeabilization, cells were washed in dPBS and incubated inLI-COR Blocking Buffer (LI-COR Biosciences, Cat #927-40000) for 1 houror longer. Plates were then incubated with an antibody specific for theMEK-dependent ERK1/2 phosphorylation sites, threonine 202 and tyrosine204 (Cell Signaling Technologies; Cat #9101), downstream of MEK in theMAP kinase signal transduction pathway, as well as GAPDH (Millipore; Cat#MAB374). pErk1/2 (Thr202/Tyr204) antibody was diluted in LI-CORblocking buffer containing 0.05% Tween-20 at 1:250; GAPDH was diluted at1:2,500. The plates were incubated overnight at 4° C. After washing withPBS/0.05% Tween-20, the cells were incubated with fluorescently-labeledsecondary antibodies (Anti-rabbit-Alexa Flour680, Invitrogen Cat#A21109; Anti-mouse-IRDye800CW, LI-COR Biosciences Cat #926-32210, bothat 1:1000 dilution) for 1 hour. Cells were then washed, as above, andanalyzed for fluorescence at both 680 and 800 nm wavelengths using theOdyssey CLx Infrared Imaging System (LI-COR Biosciences). PhosphorylatedErk1/2 (Thr202/Tyr2O4) signal was normalized to GAPDH signal for eachwell. IC₅₀ values were calculated from the normalized values using a4-parameter fit in BioAssay software and are provided in Table A.

TABLE A A375 cell IC₅₀ Ex. No. (nM)  1 562  2 1034  3 456  4 74  5 20  68  7 23  8 1  9 3 10 58 11 199 12 1060 13 178 14 15 15 17 16 274 17 25618 39 19 29 20 632 21 10 22 21 23 26 24 466 25 3 27 142 28 26 29 17 30421 31 1 32 10 33 19 34 207 35 229 36 30 37 116 38 7 39 62 40 4 41 74 42143 43 155 44 13 45 20 46 30 47 97 48 6 49 2 50 12 51 26 52 30 53 30 54167 55 28 56 56 57 94 58 45 59 72 60 508 61 11 62 17 63 114 64 2 65 2 6627 67 8 68 3 69 2

Example B MDR1 LLC-PK1 and BCRP MDCKII Permeability Assay

Both LLC-PK1 and MDR1 transfected LLC-PK1 cells were cultured and platedaccording to manufacturer's recommendations with the exception that thepassage media contained only 2% fetal bovine serum to extend passagetime out to seven days.

BCRP transfected MDCKII canine P-gp knockout cell line was cultured andplated according to manufacturer's recommendations.

Both positive and negative controls were used to assess functionality ofP-gp or BCRP efflux in the assays. Stock solutions for assay controlsand the test article were prepared in DMSO for final test concentrationsof 10 and 1 μM, respectively. Final organic concentration in the assaywas 1%. All dosing solutions contained 10 μM lucifer yellow to monitorLLC-PK1 or MDCKII cell monolayer integrity.

For the apical to basolateral determination (A to B), 75 μL of the testarticle in transport buffer were added to the apical side of theindividual transwells and 250 μL of basolateral media, without compoundor lucifer yellow, were added to each well. For the basolateral toapical determination (B to A), 250 μL of test article in transportbuffer were added to each well and 75 μL transport buffer, withoutcompound or lucifer yellow, were added to each transwell. All tests wereperformed in triplicate, and each compound was tested for both apical tobasolateral and basolateral to apical transport. The plates wereincubated for 2 hours on a Lab-Line Instruments Titer Orbital Shaker(VWR, West Chester, PA) at 50 rpm and 37° C. with 5% CO₂. All cultureplates were removed from the incubator and 50 μL of media were removedfrom the apical and basolateral portion of each well and added to 150 μLof 1 μM labetalol in 2:1 acetonitrile (acetonitrile):H₂O, v/v.

The plates were read using a Molecular Devices (Sunnyvale, CA) GeminiFluorometer to evaluate the lucifer yellow concentrations atexcitation/emission wavelengths of 425/535 nm. These values wereaccepted when found to be below 2% for apical to basolateral and 5%basolateral to apical flux across the MDR1-transfected LLC-PK1 orBCRP-transfected MDCKII cell monolayers. The plates were sealed and thecontents of each well analyzed by LC-MS/MS. The compound concentrationswere determined from the ratio of the peak areas of the compound to theinternal standard (labetalol) in comparison to the dosing solution.

LC-MS Analysis

The LC-MS/MS system was comprised of an HTS-PAL autosampler (LeapTechnologies, Carrboro, NC), an HP1200 HPLC (Agilent, Palo Alto, CA),and a MDS Sciex 4000 Q Trap system (Applied Biosystems, Foster City,CA). Chromatographic separation of the analyte and internal standard wasachieved at room temperature using a C18 column (Kinetics®, 50×300 mm,2.6 μm particle size, Phenomenex, Torrance, CA) in conjunction withgradient conditions using mobile phases A (water containing 1% isopropylalcohol and 0.1% formic acid) and B (0.1% formic acid in acetonitrile).The total run time, including re-equilibration, for a single injectionwas 1.2 minutes. Mass spectrometric detection of the analytes wasaccomplished using the ion spray positive mode. Analyte responses weremeasured by multiple reaction monitoring (MRM) of transitions unique toeach compound (the protonated precursor ion and selected product ionsfor each test article and m/z 329 to m/z 162 for labetalol, the internalstandard).

The permeability coefficient (P_(app)) is calculated from the followingequation:

P _(app)=[((C _(d) *V*(1×10⁶))/(t*0.12 cm² *C)]

where C_(d), V, t and C₀ are the detected concentration (μM), the volumeon the dosing side (mL), the incubation time (s) and the initial dosingconcentration (μM), respectively. The calculations for P_(app) were madefor each replicate and then averaged. Permeability coefficients forcompounds of Formula I are provided in Table B1. In this assay, acompound is defined has having high permeability if the permeability isgreater than 8×10⁻⁶ cm/sec, a compound is defined has having mediumpermeability if the permeability is from 2×10⁻⁶ cm/sec to 8×10⁻⁶ cm/sec,and a compound is defined has having low permeability if thepermeability is less than 2×10⁻⁶ cm/sec.

An efflux ratio is calculated from the mean apical to basolateral (A-B)P_(app) data and basolateral to apical (B-A) Papp data:

Efflux ratio=P _(app)(B-A)/P _(app)(A-B)

TABLE B1 Permeability Ex. No. (*10⁻⁶ cm/s)  1 27.2  3 46.2  4 26.1  527.8  6 32.1  7 34.5  8 34.2  9 27.1 10 26.5 14 31.1 15 27.5 19 25.9 2128.8 22 24.6 23 30.4 25 26.4 27 25.1 28 26.1 29 32.8 31 24.8 32 24.8 3324.7 36 31.1 37 28.6 38 24.6 39 32.3 40 28.4 42 29.4 44 33.0 45 31.8 4637.0 47 31.5 48 16.2 49 39.3 50 36.7 51 41.2 52 36.8 53 36.9 55 37.4 5632.1 57 31.8 58 N/A 59 N/A 60 N/A 61 26.9 62 26.1 63 N/A 64 N/A 65 12.566 24.9 67 28.3 68 18.7 69 19.6 N/A: Not available

An efflux ratio is calculated from the mean apical to basolateral (A-B)P_(app) data and basolateral to apical (B-A) Papp data:

Efflux ratio=P _(app)(B-A)/P _(app)(A-B)

Table B2 provides efflux ratios for compounds of Formula I when testedin this assay.

TABLE B2 LLC-PK1 MDCKII-hBCRP knock-in, Ex. No. MDR1 canine pg-P KO  11.4 1.1  3 1.0 0.9  4 1.3 1.1  5 1.6 1.0  6 3.4 5.5  7 5.4 9.6  8 7.13.4  9 7.7 3.2 10 6.7 6.3 14 2.7 3.4 15 3.7 2.2 18 2.3 2.0 19 3.3 1.5 214.2 3.0 22 6.2 9.0 23 3.2 2.0 25 5.7 14.1 27 0.9 1.4 28 1.1 1.1 29 1.51.1 31 5.3 4.4 32 6.5 5.2 33 4.0 8.3 36 2.9 3.5 37 2.7 3.3 38 4.5 2.9 393.9 3.2 40 3.6 1.9 42 2.8 3.3 44 1.1 2.7 45 2.2 1.3 46 1.1 1.2 47 1.3N/A 48 1.4 1.0 49 1.4 1.0 50 1.2 1.3 51 1.2 1.2 52 1.3 1.2 53 1.1 1.8 551.4 1.4 56 0.8 1.1 57 0.9 1.2 58 N/A N/A 59 N/A N/A 60 N/A N/A 61 3 N/A62 5 N/A 63 N/A N/A 64 N/A 1.5 65 N/A 0.8 66 N/A 1 67 N/A 1.1 68 N/A 2.569 N/A 2.3 N/A: Not available

Example C

PK (Free Brain-to-Free Plasma Ratio) (Mouse)

The ability of representative compounds to penetrate the BBB in mice wasdetermined by evaluating the unbound brain-to-unbound plasma (alsoreferred to as free brain-to-free plasma) concentration ratio in maleCD-1 mice.

Brain compound levels were generated from oral mouse PK dosing withtypical sampling times of 2, 4, 8, 12 and 24 hours post oral gavagedosing at 10 mg/kg. Brain samples were stored at −20±5° C. prior toanalysis. Concentrations of test compound in mouse brain homogenate weredetermined by liquid chromatography tandem mass spectrometry (LC-MS/MS)following protein precipitation with acetonitrile. A 12-pointcalibration curve, ranging from 0.5 to 10,000 ng/mL, was prepared induplicate. A solution of 400 μg/mL of test compound in dimethylsulfoxide (DMSO) was serially diluted (3-fold) in 100% DMSO, and then2.5 μL of each standard solution was added to 100 μL of naïve male CD-1mice brain homogenate. To mimic the extraction in the standard curve,2.5 μL of DMSO was added to all test samples. Both calibration and testbrain homogenate samples were spiked with 10 μL of an IS (1 μg/mL of astructural analog). Brain homogenate was generated by adding 0.75 mL of4:1 water:MeOH to each brain sample followed by homogenization for 1minute with bead beater tubes a 6 m/s using an MP Fast Prep-24@.Proteins were precipitated from 100 μL of brain homogenate sample by theaddition of 300 μL of acetonitrile. Samples were vortex-mixed for 5minutes and spun in an Allegra X-12R centrifuge (Beckman Coulter,Fullerton, CA; SX4750A rotor) for 15 min at about 1,500×g at 4° C. A 100μL aliquot of each supernatant was transferred via a 550 μL PersonalPipettor (Apricot Designs, Monrovia, CA) to 96-well plates and diluted1:1 with HPLC grade water. The resulting plates were sealed withaluminum for LC-MS/MS analysis.

Brain-to-plasma ratios were calculated using the concentration ofcompound measured in the brain divided by the concentration of compoundmeasured in the plasma. Brain-to-plasma ratios were always generatedfrom a single animal and time point. Free brain-to-free plasma ratioswere calculated by multiplying the brain-to-plasma ratio by the in vitrobrain homogenate free fraction divided by the in vitro plasma freefraction using the following equation: (B/P)*(B_(fu)/P_(fu)).

Table C provides free brain-to-free plasma ratios of the representativecompounds of Examples 6, 7, 8, 14, 22, 36 and 46 disclosed herein.

TABLE C Ex. # B/P ratio (free)  6 0.88-1.26  7 0.62-1.06  8 1.01-16.1 140.57-1.58 22 0.33-2.92 36 0.83-2.09 46 3.29-6.75

Synthetic Examples Intermediate 1

Methyl (E)-2,6-dichloro-4-(2-(dimethylamino)vinyl)nicotinate

Step 1. Preparation of methyl 2,6-dichloro-4-methylnicotinate. To asolution of 2,6-dichloro-4-methylnicotinic acid (1.0 g, 4.9 mmol) in 1:1MeOH:dioxane (10 mL) at 0° C. was added (trimethylsilyl)diazomethane(3.3 mL, 2M in hexanes, 6.6 mmol). The mixture was removed from the icebath and stirred for 10 minutes, then concentrated to half volume andpartitioned between water (20 mL) and EtOAc (20 mL). The aqueous layerwas extracted with EtOAc (2×20 mL) and the combined organic phases werewashed with brine (10 mL), dried over Na₂SO₄, filtered and carefullyconcentrated. The residue was purified by column chromatography elutingwith 0-15% EtOAc/petroleum ether to afford methyl2,6-dichloro-4-methylnicotinate (0.81 g, 76%). ¹H NMR (400 MHz, CDCl₃) δ7.2 (s, 1H), 4.0 (s, 3H), 2.3 (s, 3H) ppm.

Step 2. Preparation of methyl(E)-2,6-dichloro-4-(2-(dimethylamino)vinyl)nicotinate. To a solution ofmethyl 2,6-dichloro-4-methylnicotinate (810 mg, 3.68 mmol) in DMF (5 mL)was added N,N-dimethylformamide dimethylacetal (978 mL, 7.36 mmol) andthe mixture was stirred at 100° C. for 16 hours. The cooled mixture wastreated with water (40 mL), stirred for 10 minutes and then the solidswere collected by filtration, washed with water and dried in vacuo toafford methyl (E)-2,6-dichloro-4-(2-(dimethylamino)vinyl)nicotinate (764mg, 75%). ¹H NMR (400 MHz, CDCl₃) δ 7.3 (s, 1H), 7.1 (d, 1H), 4.8 (d,1H), 3.1 (s, 3H), 2.9 (s, 6H) ppm.

Intermediate 2

Methyl 4-bromo-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate

Step 1. Preparation of 4-bromo-2,6-dichloronicotinic acid. A solution of4-bromo-2,6-dichloropyridine (100 g, 440.7 mmol) in THF (1000 mL) wascooled to −78° C. LDA (2 M in THF, 242.4 mL, 484.8 mmol) was addeddropwise at −78° C. and stirring continued for 1 hour at −78° C. SolidCO₂ (155.1 g, 3.53 mol) was added portion-wise to the reaction andstirring continued for 2 hours at −78° C. The reaction was quenched byadding 1M Na₂CO₃ (1600 mL) followed by water (500 mL) and stirred for 10minutes. The aqueous layer was extracted with EtOAc (300 mL). The pHvalue of the aqueous was adjusted with 2N HCl to afford a solution withpH 2. The aqueous was then extracted with EtOAc (3×300 mL). The combinedorganic layers were dried over Na₂SO₄, filtered and concentrated toafford 4-bromo-2,6-dichloronicotinic acid (540 g, 75%). ¹H NMR (400 MHz,DMSO-ds) δ 15.51-12.87 (m, 1H), 8.13 (s, 1H) ppm.

Step 2. Preparation of4-bromo-2-chloro-6-oxo-1,6-dihydropyridine-3-carboxylic acid. A solutionof NaOH (4 M, 1.62 L, 6.46 mol) was heated to 110° C., and4-bromo-2,6-dichloronicotinic acid (70 g, 258.4 mmol) was added in oneportion. The mixture was stirred for 8 hours then cooled to 0° C. Thereaction was adjusted pH to 1 with HCl (6 M) and stirred for 30 minutes.Solids were collected by filtration and dried in vacuo to afford4-bromo-2-chloro-6-oxo-1,6-dihydropyridine-3-carboxylic acid (assumed100%).

Step 3. Preparation of methyl4-bromo-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate. To amixture of 4-bromo-2-chloro-6-oxo-1,6-dihydropyridine-3-carboxylic acid(100 g, 396 mmol) in DMF (800 mL) was added methyl iodide (168.6 g, 1.19mol, 73.98 mL) and K₂CO₃ (164.2 g, 1.19 mol) in one portion. The mixturewas stirred at 25° C. for 3 hours then poured into saturated NH₄Cl (1800mL), and the aqueous phase was extracted with EtOAc (2×500 mL). Thecombined organic phases were washed with brine (400 mL), dried overNa₂SO₄, filtered and concentrated. The combined residues (5 batches)were purified by column chromatography eluting with 2-100%EtOAc/petroleum ether to afford methyl4-bromo-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate (from5 batches, 141.83 g, 24.7%). ¹H NMR (400 MHz, CDCl₃) δ 6.86 (s, 1H),3.94 (s, 3H), 3.69-3.66 (m, 3H); MS (apci, m/z)=280.0, 282.0 (M+H).

Intermediate 3

Methyl(Z)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate

Methyl 4-bromo-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(20.0 g, 71.3 mmol,methanesulfonato(2-dicyclohexylphosphino-2′,6′-di-i-propoxy-1,1′-biphenyl)(2-amino-1,1′-biphenyl-2-yl)palladium(II)(5.96 g, 7.13 mmol) and(Z)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (14.8 g,74.9 mmol) were suspended in 1,4-dioxane (700 mL) and K₂CO₃ (53.5 mL,107 mmol) (2 N aqueous) was added. The mixture was stirred at 60° C. for6 hours then at ambient temperature for 12 hours under argon atmosphere.The reaction was partitioned between water (1500 mL) and EtOAc (500 mL).The aqueous layer was extracted with EtOAc (2×400 mL) and the combinedorganic layers were washed with brine (500 mL), dried over Na₂SO₄,filtered and concentrated. The residue was purified by columnchromatography eluting with 0-40-60% EtOAc/heptanes to afford methyl(Z)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(13.3 g, 68%). ¹H NMR (400 MHz, CDCl₃) δ 7.23 (s, 1H), 6.45 (d, 1H),4.86 (d, 1H), 4.40 (q, 2H), 3.89 (s, 3H), 3.67 (s, 3H), 1.35 (t, 3H); MS(apci, m/z)=272.0 (M+H).

Intermediate 4

Methyl4-(2-((2-(tert-butoxy)ethoxy)amino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate

Step 1. Preparation of methyl(E/Z)-4-(2-((2-(tert-butoxy)ethoxy)imino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.Methyl(Z)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(0.95 g, 3.50 mmol) and O-(2-(tert-butoxy)ethyl)hydroxylaminehydrochloride (593 mg, 3.50 mmol) were combined in 1,4-dioxane (10 mL).Et₃N (487 mL, 3.50 mmol) and HCl (1.75 mL, 6.99 mmol) (4N/dioxane) wereadded. The suspension was heated to 60° C. for 1 hour then cooled andfiltered. The filtrate was concentrated to afford methyl(E/Z)-4-(2-((2-(tert-butoxy)ethoxy)imino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(assumed 100%) as a 1:1 mixture of isomers. MS (apci, m/z)=359.1 (M+H).

Step 2. Preparation of Methyl4-(2-((2-(tert-butoxy)ethoxy)amino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.To a solution of methyl(E/Z)-4-(2-((2-(tert-butoxy)ethoxy)imino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(1.25 g, 3.48 mmol) in IPA (20 mL) was added sodium cyanoborohydride(1.09 g, 17.4 mmol) followed by acetic acid (1.0 mL, 17.4 mmol). Themixture was stirred at ambient temperature for 16 hours then partitionedbetween saturated NaHCO₃ (50 mL) and EtOAc (50 mL). The aqueous layerwas extracted with EtOAc (2×30 mL) and the combined organic phases werewashed with brine (30 mL) then dried over Na₂SO₄, filtered andconcentrated. The residue was purified by column chromatography elutingwith 0-80% EtOAc/DCM to afford methyl4-(2-((2-(tert-butoxy)ethoxy)amino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(0.66 g, 53%). ¹H NMR (400 MHz, CDCl₃) δ 6.43 (s, 1H), 3.90 (s, 3H),3.78 (t, 2H), 3.68 (s, 3H), 3.50 (t, 2H), 3.10 (t, 2H), 2.71 (t, 2H),1.20 (s, 9H) ppm; MS (apci, m/z)=361.1, 363.1 (M+H).

Intermediate 5

2-Chloro-4-ethylaniline

Step 1. Preparation of N-(4-ethylphenyl)acetamide. 4-Ethylaniline (513μL, 4.13 mmol) was dissolved in DCM (10.3 mL). After addingtriethylamine (690 μL, 4.95 mmol) and cooling to 0° C., acetic anhydride(467 μL, 4.95 mmol) was added dropwise. After 30 minutes, the reactionwas quenched by addition of saturated NaHCO₃ (50 mL). The organic layerswere separated and the aqueous layer was extracted with DCM (2×25 mL).The combined organic layers were dried over Na₂SO₄, filtered, andconcentrated. The residue was triturated with hexanes and solidscollected by filtration to afford N-(4-ethylphenyl)acetamide (600 mg,89%). ¹H NMR (400 MHz, CDCl₃) δ 7.40-7.38 (d, 2H), 7.16-7.14 (d, 2H),7.07 (br s, 1H), 2.64-2.58 (q, 2H), 2.16 (s, 3H), 1.23-1.20 (t, 3H) ppm.

Step 2. Preparation of N-(2-chloro-4-ethylphenyl)acetamide.N-(4-ethylphenyl)acetamide (50 mg, 0.31 mmol) was dissolved in DMF (613μL). After adding N-chlorosuccinimide (65 mg, 0.49 mmol), the solutionwas heated to 70° C. for 6 hours, then allowed to cool to ambienttemperature over 16 hours. The reaction mixture was poured into 2N HCl(4 mL) and stirred for 15 minutes. The mixture was extracted with EtOAc(10 mL). The organic layers were washed with water (3×10 mL), dried overNa₂SO₄, filtered, and concentrated. The residue was purified by columnchromatography, eluting with 0-25% EtOAc/hexanes, to affordN-(2-chloro-4-ethylphenyl)acetamide (36 mg, 59%). ¹H NMR (400 MHz,CDCl₃) δ 8.23-8.21 (d, 1H), 7.51 (br s, 1H), 7.20-7.19 (d, 1H),7.11-7.08 (dd, 1H), 2.63-2.57 (q, 2H), 2.23 (s, 3H), 1.23-1.20 (t, 3H)ppm.

Step 3. Preparation of 2-chloro-4-ethylaniline.N-(2-chloro-4-ethylphenyl)acetamide (36 mg, 0.18 mmol) was dissolved inEtOH (0.5 mL) and 12N HCl (0.5 mL, 6.0 mmol) was added. The mixture wasstirred for 2 hours at 120° C. The reaction mixture was cooled toambient temperature, brought to pH 10 by addition of 6N NaOH, thenextracted with MTBE (3×25 mL). The combined organic layers were driedover Na₂SO₄, filtered, and concentrated to afford2-chloro-4-ethylaniline (26 mg, 92%). ¹H NMR (400 MHz, CDCl₃) δ7.09-7.08 (d, 1H), 6.91-6.88 (dd, 1H), 6.71-6.69 (d, 1H), 2.55-2.49 (q,2H), 1.20-1.16 (t, 3H) ppm. MS (apci, m/z)=156.1 (M+H).

Intermediate 6

4-((Difluoromethyl)thio)-2-fluoroaniline

LiBF₄ (196 mg, 2.10 mmol) and LiH (17.5 mg, 2.10 mmol) were combined inDMF (8.8 mL, 1.75 mmol). 4-Amino-3-fluorobenzenethiol (250 mg, 1.75mmol) was added, and the mixture was stirred for 5 minutes at ambienttemperature. (Trifluoromethyl)trimethylsilane (0.644 mL, 4.37 mmol) wasadded quickly, then the solution stirred for 10 minutes at the sametemperature. TBAF (6 mL, 1N/THF, 6.00 mmol) was added quickly, then thesolution was stirred for 10 minutes at the same temperature. Thereaction mixture was quenched with water (50 mL). The mixture wasextracted with EtOAc (2×25 mL) and the combined organic layers werewashed with water (3×50 mL) and brine (50 mL), then dried over Na₂SO₄,filtered and concentrated. The residue was purified by columnchromatography, eluting with 0-15% EtOAc/hexanes, to afford4-((difluoromethyl)thio)-2-fluoroaniline (56 mg, 17%). ¹H NMR (400 MHz,CDCl₃) δ 7.25-7.22 (dd, 1H), 7.19-7.16 (ddd, 1H), 6.86-6.58 (t, 1H),6.77-6.73 (dd, 1H), 3.95 (br s, 1H) ppm.

Intermediate 7

2-Chloro-4-(methylthio)aniline

2-Chloro-4-iodoaniline (250 mg, 0.986 mmol), NiBr₂ (22 mg, 0.099 mmol),Zn powder (129 mg, 1.97 mmol) and 2,2′-bipyridine (15 mg, 0.099 mmol)were dissolved in THF (1.6 mL) under Ar atmosphere.1,2-Dimethyldisulfane (44 μL, 0.493 mmol) was added and the mixture wassealed and heated to 65° C. for 16 hours. The reaction mixture wasdiluted with EtOAc (50 mL) and washed with concentrated NH₄OH (50 mL)followed by 10% citric acid solution (50 mL). The combined organiclayers were dried over Na₂SO₄, filtered, and concentrated. The residuewas purified by column chromatography, eluting with 0-15% EtOAc/hexanes,to afford 2-chloro-4-(methylthio)aniline (116 mg, 68%). ¹H NMR (400 MHz,CDCl₃) δ 7.27-7.26 (m, 1H), 7.09-7.06 (dd, 1H), 6.71-6.69 (d, 1H), 4.02(br s, 2H), 2.41 (s, 3H) ppm. MS (apci, m/z)=174.0 (M+H).

Intermediate 8

Methyl(E)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate

Methyl 4-bromo-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(1.0 g, 3.565 mmol),Methanesulfonato(2-dicyclohexylphosphino-2′,6′-di-i-propoxy-1,1′-biphenyl)(2-amino-1,1′-biphenyl-2-yl)palladium(II)(0.2982 g, 0.3565 mmol), and(E)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (0.7414 g,3.743 mmol) were suspended in 1,4-dioxane (35.65 mL) and potassiumcarbonate (2.674 mL, 2N aqueous, 5.35 mmol) was added. After degassingwith argon, the mixture was stirred at 60° C. for 4 hours. The cooledreaction was partitioned between water (150 mL) and EtOAc (50 mL). Theaqueous layer was washed with EtOAc (2×40 mL). The combined organiclayers were washed with brine (150 mL), dried over sodium sulfate,filtered and concentrated. The residue was purified by columnchromatography eluting with 0-40% EtOAc/heptane to afford methyl(E)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(379 mg, 39%). ¹H NMR (400 MHz, CDCl₃) δ 7.27 (s, 1H), 7.03 (d, 1H),6.43 (s, 1H), 5.53 (d, 1H), 3.95-3.83 (m, 5H), 3.67 (s, 3H), 1.34 (t,2H) ppm. MS (apci, m/z)=272.1 (M+H).

Example 1

8-((4-bromo-2-fluorophenyl)amino)-2-cyclopropyl-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of methyl 2,6-dichloro-4-(2-oxoethyl)nicotinate. Asuspension of methyl(E)-2,6-dichloro-4-(2-(dimethylamino)vinyl)nicotinate (0.797 g, 2.90mmol) in Et₂O (30 mL) and 1 N HCl (30 mL) was stirred vigorously atambient temperature for 1 hour. The resulting solution was treated withbrine (20 mL) and extracted with Et₂O (3×10 mL). The combined organicphases were washed with brine (10 mL), dried over MgSO₄, filtered andcarefully concentrated to afford methyl2,6-dichloro-4-(2-oxoethyl)nicotinate (assumed 100%) which was usedimmediately.

Step 2. Preparation of8-chloro-2-cyclopropyl-6-methoxy-3,4-dihydro-2,7-naphthyridin-1(2H)-one.To a solution of methyl 2,6-dichloro-4-(2-oxoethyl)nicotinate (0.719 g,2.90 mmol) in 1:1 IPA:MeOH (20 mL) at 0° C. was added cyclopropylamine(201 mL, 2.90 mmol). The mixture was stirred at ambient temperature for10 minutes then sodium cyanoborohydride (546 mg, 8.70 mmol) and aceticacid (498 mL, 8.70 mmol) were added. After stirring at ambienttemperature for 16 hours, the mixture was partitioned between saturatedNaHCO₃ (50 mL) and EtOAc (20 mL) and the aqueous layer was extractedwith EtOAc (2×20 mL). The combined organic phases were washed with brine(10 mL), dried over Na₂SO₄, filtered and concentrated. The residue wasdissolved in methanol (10 mL), treated with 1N NaOH (3.33 mL, 3.33 mmol)and stirred at ambient temperature for 1 hour. The mixture wasconcentrated to half volume then partitioned between water (20 mL) andDCM (20 mL). The aqueous layer was extracted with DCM (2×10 mL) and thecombined organic phases were washed with brine (10 mL), dried overNa₂SO₄, filtered and concentrated. The residue was purified by columnchromatography eluting with 0-10% (20% MeOH/DCM)/DCM to afford8-chloro-2-cyclopropyl-6-methoxy-3,4-dihydro-2,7-naphthyridin-1(2H)-one(310 mg, 42%). ¹H NMR (400 MHz, CDCl₃) δ 6.5 (s, 1H), 4.0 (s, 3H), 3.5(t, 2H), 2.9 (m, 3H), 0.9 (m, 2H), 0.7 (m, 2H) ppm; MS (apci,m/z)=253.1, 255.1 (M+H).

Step 3. Preparation of8-chloro-2-cyclopropyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of8-chloro-2-cyclopropyl-6-methoxy-3,4-dihydro-2,7-naphthyridin-1(2H)-one(99 mg, 0.39 mmol) in acetonitrile (2 mL) was added trimethylsilyliodide (2.94 mL, 1.96 mmol). The mixture was stirred at ambienttemperature for 48 hours then concentrated. The residue was purified bycolumn chromatography eluting with 0-15-20% (20% MeOH/DCM)/DCM to afford8-chloro-2-cyclopropyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(assume 100%) which was used immediately. MS (apci, m/z)=239.0 (M+H).

Step 4. Preparation of8-chloro-2-cyclopropyl-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of8-chloro-2-cyclopropyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(93 mg, 0.39 mmol) in THF (2 mL) was added K₂CO₃ (110 mg, 0.78 mmol)followed by methyl iodide (29 mL, 0.47 mmol). The mixture was stirred atambient temperature for 2 hours then treated with additional methyliodide (50 mL, 0.84 mmol) and stirred for a further 16 hours. Themixture was filtered, washed with (4:1) DCM/MeOH (10 mL) and thefiltrate concentrated. The residue was purified by column chromatographyeluting with 0-15% (20% MeOH/DCM)/DCM to afford8-chloro-2-cyclopropyl-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(31 mg, 31%). ¹H NMR (400 MHz, CDCl₃) δ 6.29 (s, 1H), 3.73 (s, 3H), 3.45(t, 2H), 2.87-2.81 (m, 1H), 2.77 (t, 2H), 0.94-0.89 (m, 2H), 0.72-0.68(m, 2H) ppm; MS (apci, m/z)=253.1 (M+H).

Step 5. Preparation of8-((4-bromo-2-fluorophenyl)amino)-2-cyclopropyl-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of 4-bromo-2-fluoroaniline (24 mg, 0.12 mmol) in THF (1mL) at −78° C. under N₂ was added LiHMDS (1.84 mL, 1.0 M/THF, 0.184mmol). The mixture was stirred for 45 minutes then a suspension of8-chloro-2-cyclopropyl-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(31 mg, 0.13 mmol) in THF (2 mL) was added. The mixture was stirred at−78° C. for 20 minutes then quenched with saturated NH₄Cl (10 mL),stirred at ambient temperature for 10 minutes then extracted with EtOAc(3×10 mL). The combined organic phases were washed with brine (10 mL),dried over Na₂SO₄, filtered and concentrated. The residue was purifiedby column chromatography eluting with 0-80% EtOAc/hexanes to afford8-((4-bromo-2-fluorophenyl)amino)-2-cyclopropyl-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(24.9 mg, 50%). ¹H NMR (400 MHz, CDCl₃) δ 11.6 (s, 1H), 7.29 (dd, 1H),7.19 (ddd, 1H), 6.71 (t, 1H), 5.99 (s, 1H), 3.50 (t, 2H), 3.20 (s, 3H),2.77 (t, 2H), 2.74-2.70 (m, 1H), 0.93-0.87 (m, 2H), 0.72-0.67 (m, 2H)ppm; MS (apci, m/z)=406.0, 408.0 (M+H).

Example 2

8-((4-bromo-2-fluorophenyl)amino)-2-(cyclopropylmethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 1, substitutingO-(cyclopropylmethyl)hydroxylamine hydrochloride in place ofcyclopropylamine in step 2 to afford8-((4-bromo-2-fluorophenyl)amino)-2-(cyclopropylmethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(20 mg, 57%). ¹H NMR (400 MHz, CDCl₃) δ 11.32 (s, 1H), 7.30 (dd, 1H),7.21 (m, 1H), 6.72 (t, 1H), 6.00 (s, 1H), 3.85 (d, 2H), 3.77 (t, 2H),3.18 (s, 3H), 2.98 (t, 2H), 1.22-1.14 (m, 1H), 0.64-0.59 (m, 2H),0.36-0.31 (m, 2H) ppm; MS (apci, m/z)=436.0, 438.0 (M+H).

Example 3

8-((4-bromo-2-fluorophenyl)amino)-2-ethoxy-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 1, substituting O-ethylhydroxylaminehydrochloride in place of cyclopropylamine in step 2 to afford8-((4-bromo-2-fluorophenyl)amino)-2-ethoxy-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(22 mg, 55%). ¹H NMR (400 MHz, CDCl₃) δ 11.34 (s, 1H), 7.30 (dd, 1H),7.21 (m, 1H), 6.72 (t, 1H), 5.99 (s, 1H), 4.08 (q, 2H), 3.73 (t, 2H),3.19 (s, 3H), 2.99 (t, 2H), 1.32 (t, 3H) ppm; MS (apci, m/z)=410.0,412.0 (M+H).

Example 4

2-cyclopropyl-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 1, substituting2-fluoro-4-(methylthio)aniline in place of 4-bromo-2-fluoroaniline instep 5 to afford2-cyclopropyl-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(50.4 mg, 66%). ¹H NMR (400 MHz, CDCl₃) δ 11.62 (s, 1H), 7.01 (dd, 1H),6.95 (dd, 1H), 6.79 (t, 1H), 5.95 (s, 1H), 3.49 (t, 2H), 3.18 (s, 3H),2.80-2.68 (m, 3H), 2.47 (s, 3H), 0.93-0.87 (m, 2H), 0.72-0.67 (m, 2H)ppm; MS (apci, m/z)=374.1 (M+H).

Example 5

2-cyclopropyl-8-((2-fluoro-4-iodophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 1 substituting 2-fluoro-4-iodoaniline inplace of 4-bromo-2-fluoroaniline in step 5 to afford2-cyclopropyl-8-((2-fluoro-4-iodophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(46.7 mg, 52%). ¹H NMR (400 MHz, CDCl₃) δ 11.60 (s, 1H), 7.46 (dd, 1H),7.37 (dt, 1H), 6.55 (t, 1H), 6.00 (s, 1H), 3.50 (t, 2H), 3.21 (s, 3H),2.81-2.69 (m, 3H), 0.93-0.87 (m, 2H), 0.72-0.67 (m, 2H) ppm; MS (apci,m/z)=454.0 (M+H).

Example 6

8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

8-((2-Fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionewas prepared either by Method A or Method B.

Method A.

Step 1. Preparation of 4-bromo-2,6-dichloronicotinic acid. A solution of4-bromo-2,6-dichloropyridine (100 g, 440.7 mmol) in THF (1000 mL) wascooled to −78° C. LDA (242.4 mL, 2M/THF, 484.8 mmol) was added dropwiseat −78° C. and stirring continued for 1 hour at −78° C. Solid CO₂ (155.1g, 3.53 mol) was added portion-wise to the reaction and stirringcontinued for 2 hours at −78° C. The reaction was quenched by adding 1MNa₂CO₃ (1600 mL) followed by water (500 mL) and stirred for 10 minutes.The aqueous layer was extracted with EtOAc (300 mL). The pH value of theaqueous was adjusted with 2N HCl to afford a solution with pH 2. Theaqueous was then extracted with EtOAc (3×300 mL). The combined organiclayers were dried over Na₂SO₄, filtered and concentrated to afford4-bromo-2,6-dichloronicotinic acid (540 g, 75%). ¹H NMR (400 MHz,DMSO-d₆) δ 15.51-12.87 (m, 1H), 8.13 (s, 1H) ppm.

Step 2. Preparation of4-bromo-2-chloro-6-oxo-1,6-dihydropyridine-3-carboxylic acid. A solutionof NaOH (1.62 L, 4M, 6.46 mol) was heated to 110° C., and4-bromo-2,6-dichloronicotinic acid (70 g, 258.4 mmol) obtained in step 1was added in one portion. The mixture was stirred for 8 hours thencooled to 0° C. The reaction was adjusted pH to 1 with HCl (6 M) andstirred for 30 minutes. Solids were collected by filtration and dried invacuo to afford 4-bromo-2-chloro-6-oxo-1,6-dihydropyridine-3-carboxylicacid (assumed 100%).

Step 3. Preparation of methyl4-bromo-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate. To amixture of 4-bromo-2-chloro-6-oxo-1,6-dihydropyridine-3-carboxylic acid(100 g, 396 mmol) in DMF (800 mL) was added methyl iodide (168.6 g, 1.19mol, 73.98 mL) and K₂CO₃ (164.2 g, 1.19 mol) in one portion. The mixturewas stirred at 25° C. for 3 hours then poured into saturated NH₄Cl (1800mL), and the aqueous phase was extracted with EtOAc (2×500 mL). Thecombined organic phases were washed with brine (400 mL), dried overNa₂SO₄, filtered and concentrated. The combined residues (5 batches)were purified by column chromatography eluting with 2-100%EtOAc/petroleum ether to afford methyl4-bromo-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate (from5 batches, 141.83 g, 24.7%). ¹H NMR (400 MHz, CDCl₃) δ 6.86 (s, 1H),3.94 (s, 3H), 3.69-3.66 (m, 3H); MS (apci, m/z)=280.0, 282.0 (M+H).

Step 4. Preparation of methyl(Z)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.Methyl 4-bromo-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(20.0 g, 71.3 mmol) obtained in step 3,methanesulfonato(2-dicyclohexylphosphino-2′,6′-di-i-propoxy-1,1′-biphenyl)(2-amino-1,1′-biphenyl-2-yl)palladium(II)(5.96 g, 7.13 mmol) and(Z)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (14.8 g,74.9 mmol) were suspended in 1,4-dioxane (700 mL) and K₂CO₃ (53.5 mL, 2Naq, 107 mmol) was added. The mixture was stirred at 60° C. for 6 hoursthen at ambient temperature for 12 hours under argon atmosphere. Thereaction was partitioned between water (1500 mL) and EtOAc (500 mL). Theaqueous layer was extracted with EtOAc (2×400 mL) and the combinedorganic layers were washed with brine (500 mL), dried over Na₂SO₄,filtered and concentrated. The residue was purified by columnchromatography eluting with 0-40-60% EtOAc/heptanes to afford methyl(Z)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(13.3 g, 68%). ¹H NMR (400 MHz, CDCl₃) δ 7.23 (s, 1H), 6.45 (d, 1H),4.86 (d, 1H), 4.40 (q, 2H), 3.89 (s, 3H), 3.67 (s, 3H), 1.35 (t, 3H)ppm; MS (apci, m/z)=272.0 (M+H).

Step 5. Preparation of methyl(E/Z)-4-(2-((2-(tert-butoxy)ethoxy)imino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.Methyl(Z)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(0.95 g, 3.50 mmol) obtained in step 4 andO-(2-(tert-butoxy)ethyl)hydroxylamine hydrochloride (593 mg, 3.50 mmol)were combined in 1,4-dioxane (10 mL). Et₃N (487 mL, 3.50 mmol) and HCl(1.75 mL, 4N/dioxane, 6.99 mmol) were added. The suspension was heatedto 60° C. for 1 hour then cooled and filtered. The filtrate wasconcentrated to afford methyl(E/Z)-4-(2-((2-(tert-butoxy)ethoxy)imino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(assumed 100%) as a 1:1 mixture of isomers. MS (apci, m/z)=359.1 (M+H).

Step 6. Preparation of Methyl4-(2-((2-(tert-butoxy)ethoxy)amino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.To a solution of methyl(E/Z)-4-(2-((2-(tert-butoxy)ethoxy)imino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(1.25 g, 3.48 mmol) in IPA (20 mL) obtained in step 5 was added sodiumcyanoborohydride (1.09 g, 17.4 mmol) followed by acetic acid (1.0 mL,17.4 mmol). The mixture was stirred at ambient temperature for 16 hoursthen partitioned between saturated NaHCO₃ (50 mL) and EtOAc (50 mL). Theaqueous layer was extracted with EtOAc (2×30 mL) and the combinedorganic phases were washed with brine (30 mL) then dried over Na₂SO₄,filtered and concentrated. The residue was purified by columnchromatography eluting with 0-80% EtOAc/DCM to afford methyl4-(2-((2-(tert-butoxy)ethoxy)amino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(0.66 g, 53%). ¹H NMR (400 MHz, CDCl₃) δ 6.43 (s, 1H), 3.90 (s, 3H),3.78 (t, 2H), 3.68 (s, 3H), 3.50 (t, 2H), 3.10 (t, 2H), 2.71 (t, 2H),1.20 (s, 9H) ppm; MS (apci, m/z)=361.1, 363.1 (M+H).

Step 7. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.2-Fluoro-4-(methylthio)aniline (114 mg, 0.73 mmol) was dissolved in THF(5.0 mL, 0.69 mmol) and cooled to −78° C. under N₂ atmosphere. LiHMDS(1.39 mL, 1.0 m/THF, 1.39 mmol) was added dropwise, then the mixturestirred for 45 minutes. A solution of methyl4-(2-((2-(tert-butoxy)ethoxy)amino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(250 mg, 0.69 mmol) obtained in step 6 in THF (5.0 mL) was then addeddropwise. After stirring for 10 minutes, additional LiHMDS (1.0 mL, 1.0mmol) was added and stirring continued at −78° C. for 1 hour. Thereaction mixture was quenched with saturated NH₄Cl (20 mL), thenextracted with EtOAc (3×10 mL). The combined organic phases were driedover Na₂SO₄, filtered and concentrated. The residue was purified bycolumn chromatography eluting with 0-15% (20% MeOH/DCM)/DCM) to afford2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(111 mg, 36%). ¹H NMR (400 MHz, CDCl3) δ 11.35 (s, 1H), 7.02-6.99 (dd,1H), 6.97-6.94 (m, 1H), 6.82-6.77 (t, 1H), 5.95 (s, 1H), 4.16-4.14 (m,2H), 3.80-3.77 (t, 2H), 3.62-3.60 (m, 2H), 3.17 (s, 3H), 2.99-2.95 (t,2H), 2.47 (s, 3H), 1.21 (s, 9H) ppm; MS (apci, m/z)=450.2 (M+H).

Step 8. Preparation of8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(111 mg, 0.247 mmol) obtained in step 7 in acetonitrile (2.0 mL) wasadded H₃PO₄ (2.0 mL, 38.5 mmol). The solution was heated to 60° C. for15 minutes then cooled and quenched with saturated NaHCO₃ (20 mL). Themixture was extracted with EtOAc (3×10 mL) and the combined organicphases were dried over Na₂SO₄, filtered and concentrated. The residuewas purified by column chromatography eluting with 0-20% (20%MeOH/DCM)/DCM, then by reverse phase HPLC (5-95% acetonitrile/water/0.1%TFA over 30 min). Fractions containing the clean desired product werecombined and the product was converted to the free base with DCM/NaHCO₃to afford8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(62 mg, 64%). ¹H NMR (400 MHz, CDCl₃) δ 11.17 (s, 1H), 7.03-6.97 (m,2H), 6.85 (t, 1H), 5.95 (s, 1H), 4.03 (t, 2H), 3.74-3.70 (m, 4H), 3.16(s, 3H), 2.98 (t, 2H), 2.48 (s, 3H) ppm; MS (apci, m/z)=394.1 (M+H).PXRD analysis was performed as described in Example 77. The PXRDpattern, which is shown in FIG. 4 , confirmed that the material obtainedis amorphous. On this basis, the material was designated as amorphous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 4.

Method B.

Step 1. Preparation of methyl4-bromo-2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.To a solution of methyl4-bromo-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(prepared according to any of the methods described herein; 50.0 g,87.2%, 155 mmol) and 2-fluoro-4-(methylthio)aniline (24.4 g, 55 mmol) inTHF (50 mL) at 0° C. was added potassium tert-butoxide (34.9 g, 311 mL,1.0 molar, 311 mmol) over 40 minutes. The reaction was held at 0° C. for50 minutes then diluted with saturated NH₄Cl (600 mL) and EtOAc (500mL). Water was added to dissolve solids and the layers were separated.The aqueous layer was extracted with EtOAc (2×300 mL) and the combinedorganic phases were washed with water (500 mL) and brine (500 mL), thendried over Na₂SO₄, filtered and concentrated. The residue was suspendedin MeOH (57 mL) and briefly sonicated then heated to 60° C. for 10minutes. The mixture was allowed to cool slowly to ambient temperature,then cooled to 0° C. for 30 minutes. Solids were filtered and collectedto afford methyl4-bromo-2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(40.14 g, 64.4%). ¹H NMR CDCl₃ δ 9.03 (s, 1H), 7.04 (dd, 1H), 6.96 (m,1H), 6.71-6.65 (m, 2H), 3.85 (s, 3H), 3.24 (s, 3H), 2.47 (s, 3H) ppm. MS(apci, m/z)=401.0, 403.0 (M+H).

Step 2. Preparation of methyl(Z)-4-(2-ethoxyvinyl)-2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.(Z)-2-(2-Ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (80.25 g,82.3 mL, 405.17 mmol) was added to a stirred solution of methyl4-bromo-2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(130.06 g, 324.14 mmol) obtained according to the procedure of step 1,K₂CO₃ (67.2 g, 243.10 mL, 2M, 486.21 mmol) andmethanesulfonato(2-dicyclohexylphosphino-2′,6′-di-i-′propoxy-1,1′-biphenyl)′2-amino-1,1′-biphenyl-2-yl)palladium(II)(6.78 g, 8.10 mmol) in 2-methyltetrahydrofuran (860 mL) at ambienttemperature under argon. The reaction mixture was sparged with argon for30 minutes. The reaction was heated to 60° C. and stirred for 13 hours,then stirred at ambient temperature for 6 hours. The reaction wasdiluted with water (1800 mL) and EtOAc (1100 mL), then filtered throughGF/F paper. Filtrate layers were separated, and the aqueous layer wasextracted with EtOAc (2×650 mL). The combined organic layers were washedwith brine (1500 mL), dried over Na₂SO₄, filtered and concentrated. Theresidue was treated with MTBE (30 mL), concentrated and dried in vacuo.The residue was suspended in MTBE (330 mL) and stirred at 60° C. for 1hour. The mixture was allowed to cool slowly to ambient temperature thenstirred at 0° C. for 40 minutes. Solids were filtered, washed with coldMTBE (110 mL), then cold MTBE:heptane (1:1 mixture, 150 mL) and dried invacuo to afford methyl(Z)-4-(2-ethoxyvinyl)-2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(98.54 g, 77.46%). 1H NMR CDCl₃ 8.65 (s, 1H), 7.04 (dd, 1H), 6.97 (s,1H), 6.92 (m, 1H), 6.56 (t, 1H), 6.37 (d, 1H), 5.36 (d, 1H), 4.01 (q,2H), 3.79 (s, 3H), 3.31 (s, 3H), 2.46 (s, 3H), 1.35 (t, 3H) ppm. MS(apci, m/z)=393.1 (M+H).

Step 3. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Triethylamine (2.84 g, 3.91 mL, 28.0 mmol) was added to a stirredsolution of methyl(Z)-4-(2-ethoxyvinyl)-2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(10.0 g, 25.5 mmol) obtained according to the procedure of step 2 andO-(2-(tert-butoxy)ethyl)hydroxylamine hydrochloride (4.95 g, 96%, 28.0mmol) in 1,4-dioxane (255 mL). The reaction was placed in a bath of coldwater, purged with Ar, and HCl (2.04 g, 14.0 mL, 4M/dioxanes, 56.1 mmol)was added dropwise via addition funnel. The mixture was heated at 60° C.for 1 hour, then cooled to ambient temperature using a cold water bath.Pyridine borane (4.74 g, 5.42 mL, 51.0 mmol) was slowly added and themixture stirred for 10 minutes at ambient temperature. HCl (1.11 g, 7.64mL, 4M/dioxane, 30.6 mmol) was added over 5 minutes, and stirringcontinued for 20 minutes. The reaction was heated at 60° C. for 21hours, then cooled to ambient temperature. The cooled mixture wasneutralized to about pH 7 by slow addition of saturated NaHCO₃ (140 mL),then diluted with EtOAc (400 mL) and H₂O (300 mL). The layers wereseparated, and the aqueous layer was extracted with EtOAc (2×150 mL).The combined organic phases were washed with brine (600 mL), dried overNa₂SO₄, filtered and concentrated. The residue was dissolved in minimalDCM and filtered through a 5 inch silica plug, eluting with 3% MeOH:DCM(1 L). The filtrate was concentrated to afford2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(assumed 100%) which was used without further purification. MS (apci,m/z)=450.2 (M+H).

Step 4. Preparation of8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(14.18 g, 31.54 mmol) obtained according to the procedure of step 3 inacetonitrile (63.09 mL, 31.54 mmol) at ambient temperature was addedphosphoric acid (36.36 g, 21.60 mL, 85%, 315.4 mmol) dropwise, over 20minutes. The reaction was heated to 60° C. for 4 hours, then cooledslowly to ambient temperature. The reaction was poured slowly into acold solution of potassium phosphate (66.95 g, 157.7 mL, 2M, 315.4 mmol)in an ice bath, with vigorous stirring. After stirring for 10 minutes,EtOAc (300 mL) was added and the layers were separated. The organiclayer was washed with brine (300 mL), then the aqueous layer extractedwith EtOAc (100 mL). The combined organic phases were dried over Na₂SO₄,filtered and concentrated. The residue was purified by columnchromatography, eluting with 0-40% (20% MeOH/DCM)/MTBE to afford8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(9.48 g, 76.4%). PXRD analysis confirmed that the material obtained isamorphous ¹H NMR CDCl₃ δ 11.19 (s, 1H), 7.06-6.93 (m, 2H), 6.91-6.76 (m,1H), 5.95 (t, 1H), 4.44 (s, 1H), 4.21-3.87 (m, 2H), 3.84-3.54 (m, 4H),3.16 (s, 3H), 2.98 (td, 2H), 2.48 (s, 3H). MS (apci, m/z)=394.1 (M+H).

Example 7

8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 6, Method A, substituting4-bromo-2-fluoroaniline in place of 2-fluoro-4-(methylthio)aniline instep 7 to afford8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(15 mg, 63%). ¹H NMR (400 MHz, CDCl₃) δ 11.14 (s, 1H), 7.33 (dd, 1H),7.24 (m, 1H), 6.78 (t, 1H), 5.99 (s, 1H), 4.38 (t, 1H), 4.05-4.01 (m,2H), 3.75-3.70 (m, 4H), 3.17 (s, 3H), 3.00 (t, 2H) ppm; MS (apci,m/z)=426.0, 428.0 (M+H).

Example 8

8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 6, Method A, substituting4-iodo-2-fluoroaniline in place of 2-fluoro-4-(methylthio)aniline instep 7 to afford8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(12 mg, 74%). ¹H NMR (400 MHz, CDCl₃) δ 11.11 (s, 1H), 7.49 (dd, 1H),7.42 (m, 1H), 6.63 (t, 1H), 6.00 (s, 1H), 4.42-4.33 (m, 1H), 4.05-4.00(m, 2H), 3.76-3.69 (m, 4H), 3.18 (s, 3H), 2.99 (t, 2H) ppm; MS (apci,m/z)=474.0 (M+H).

Example 9

8-((2-chloro-4-iodophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 6, Method A, substituting2-chloro-4-iodoaniline in place of 2-fluoro-4-(methylthio)aniline instep 7 to afford8-((2-chloro-4-iodophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(6.4 mg, 50%). ¹H NMR (400 MHz, CDCl₃) δ 11.09 (s, 1H), 7.79 (s, 1H),7.48 (d, 1H), 6.46 (d, 1H), 6.02 (s, 1H), 4.03 (t, 2H), 3.78-3.70 (m,4H), 3.14 (s, 3H), 3.00 (t, 2H) ppm; MS (apci, m/z)=490.0 (M+H).

Example 10

8-((4-bromo-2-chlorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 6, Method A, substituting4-bromo-2-chloroaniline in place of 2-fluoro-4-(methylthio)aniline instep 7 to afford8-((4-bromo-2-chlorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(7.9 mg, 52%). ¹H NMR 11.10 (s, 1H), 7.62 (d, 1H), 7.31 (dd, 1H), 6.61(d, 1H), 6.02 (s, 1H), 4.03 (t, 2H), 3.77-3.68 (m, 4H), 3.14 (s, 3H),3.00 (t, 2H) ppm; MS (apci, m/z)=442.0, 444.0 (M+H).

Example 11

8-((4-bromo-2,3-difluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 6, Method A, substituting4-bromo-2,3-difluoroaniline in place of 2-fluoro-4-(methylthio)anilinein step 7 to afford8-((4-bromo-2,3-difluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(12.2 mg, 75%). ¹H NMR 11.17 (s, 1H), 7.30-7.22 (m, 1H), 6.65 (dt, 1H),6.04 (s, 1H), 4.32 (bs, 1H), 4.04 (t, 2H), 3.78-3.69 (m, 4H), 3.21 (s,3H), 3.01 (t, 2H) ppm; MS (apci, m/z)=444.0 (M+H).

Example 12

8-((4-bromo-3-chloro-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 6, Method A, substituting4-bromo-3-chloro-2-fluoroaniline in place of2-fluoro-4-(methylthio)aniline in step 7 to afford8-((4-bromo-3-chloro-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(10.6 mg, 68%). ¹H NMR 11.17 (s, 1H), 7.37 (d, 1H), 6.68 (t, 1H), 6.04(s, 1H), 4.06-4.00 (m, 2H), 3.77-3.69 (m, 4H), 3.20 (s, 3H), 3.01 (t,2H) ppm; MS (apci, m/z)=460.0, 462.0 (M+H).

Example 13

8-((2-fluoro-4-(trifluoromethyl)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(trifluoromethyl)Phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 7 of Method A, substituting2-fluoro-4-(trifluoromethyl)aniline in place of2-fluoro-4-(methylthio)aniline to afford2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(trifluoromethyl)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(11 mg, 22%). MS (apci, m/z)=472.2 (M+H).

Step 2. Preparation of8-((2-fluoro-4-(trifluoromethyl)Phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 8 of Method A, substituting2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(trifluoromethyl)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionein place of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dioneto afford8-((2-fluoro-4-(trifluoromethyl)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(6 mg, 62%) H NMR (400 MHz, CDCl₃) δ 11.18 (s, 1H), 7.44-7.41 (dd, 1H),7.38-7.36 (d, 1H), 6.92-6.88 (t, 1H), 6.08 (s, 1H), 4.05-4.03 (m, 2H),3.76-3.72 (m, 4H), 3.23 (s, 3H), 3.03-3.00 (t, 2H) ppm; MS (apci,m/z)=416.1 (M+H).

Example 14

8-((4-ethyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((4-ethyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 7 of Method A, substituting4-ethyl-2-fluoroaniline in place of 2-fluoro-4-(methylthio)aniline toafford2-(2-(tert-butoxy)ethoxy)-8-((4-ethyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(19 mg, 42%). MS (apci, m/z)=432.2 (M+H).

Step 2. Preparation of8-((4-ethyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 8 of Method A, substituting2-(2-(tert-butoxy)ethoxy)-8-((4-ethyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionein place of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dioneto afford8-((4-ethyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(10 mg, 61%). ¹H NMR (400 MHz, CDCl₃) δ 11.17 (s, 1H), 7.00-6.91 (m,2H), 6.87-6.83 (t, 1H), 5.93 (s, 1H), 4.03-4.01 (m, 2H), 3.74-3.70 (m,4H), 3.15 (s, 3H), 3.00-2.97 (t, 2H), 2.66-2.61 (q, 2H), 1.25-1.21 (t,3H) ppm; MS (apci, m/z)=376.2 (M+H).

Example 15

8-((4-cyclopropyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((4-cyclopropyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 7 of Method A, substituting4-cyclopropyl-2-fluoroaniline in place of 2-fluoro-4-(methylthio)anilineto afford2-(2-(tert-butoxy)ethoxy)-8-((4-cyclopropyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(32 mg, 52%). MS (apci, m/z)=444.2 (M+H).

Step 2. Preparation of8-((4-cyclopropyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 8 of Method A, substituting2-(2-(tert-butoxy)ethoxy)-8-((4-cyclopropyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionein place of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dioneto afford8-((4-cyclopropyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(19 mg, 68%). ¹H NMR (400 MHz, CDCl₃) δ 11.18 (s, 1H), 6.83-6.78 (m,3H), 5.92 (s, 1H), 4.49 (br s, 1H), 4.03-4.01 (m, 2H), 3.74-3.69 (m,4H), 3.13 (s, 3H), 3.0-2.96 (t, 2H), 1.90-1.84 (m, 1H), 1.03-0.98 (m,2H), 0.69-0.65 (m, 2H) ppm; MS (apci, m/z)=388.1 (M+H).

Example 16

8-((2-fluoro-4-methoxyphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-methoxyphenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 7 of Method A, substituting2-fluoro-4-methoxyaniline in place of 2-fluoro-4-(methylthio)aniline toafford2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-methoxyphenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(8 mg, 17%). MS (apci, m/z)=434.2 (M+H).

Step 2. Preparation of8-((2-fluoro-4-methoxyphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 8 of Method A, substituting2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-methoxyphenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionein place of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dioneto afford8-((2-fluoro-4-methoxyphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(3 mg, 43%). ¹H NMR (400 MHz, CDCl₃) δ 11.18 (s, 1H), 6.94-6.90 (t, 1H),6.07-6.64 (m, 2H), 5.91 (s, 1H), 4.04-4.01 (m, 2H), 3.80 (s, 3H),3.75-3.69 (m, 4H), 3.12 (s, 3H), 3.00-2.96 (t, 2H) ppm; MS (apci,m/z)=378.1 (M+H).

Example 17

8-((2-fluoro-4-((trifluoromethyl)thio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-((trifluoromethyl)thio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 7 of Method A, substituting2-fluoro-4-((trifluoromethyl)thio)aniline in place of2-fluoro-4-(methylthio)aniline to afford2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-((trifluoromethyl)thio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(17 mg, 31%). MS (apci, m/z)=504.1 (M+H).

Step 2. Preparation of8-((2-fluoro-4-((trifluoromethyl)thio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 8 of Method A, substituting2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-((trifluoromethyl)thio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionein place of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dioneto afford8-((2-fluoro-4-((trifluoromethyl)thio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(10 mg, 66%). ¹H NMR (400 MHz, CDCl₃) δ 11.16 (s, 1H), 7.49-7.46 (dd,1H), 7.40-7.38 (d, 1H), 6.86-6.82 (t, 1H), 6.06 (s, 1H), 4.31 (br s,1H), 4.05-4.02 (t, 2H), 3.76-3.72 (m, 4H), 3.23 (s, 3H), 3.03-3.00 (t,2H); MS (apci, m/z)=448.1 (M+H).

Example 18

8-((2-fluoro-4-isopropylphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-isopropylphenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 7 of Method A, substituting2-fluoro-4-isopropylaniline in place of 2-fluoro-4-(methylthio)anilineto afford2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-isopropylphenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(17 mg, 34%). MS (apci, m/z)=446.2 (M+H).

Step 2. Preparation of8-((2-fluoro-4-isopropylphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 8 of Method A, substituting2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-isopropylphenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionein place of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dioneto afford8-((2-fluoro-4-isopropylphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(10 mg, 67%). ¹H NMR (400 MHz, CDCl₃) δ 11.16 (s, 1H), 7.00-6.93 (m,2H), 6.97-6.83 (t, 1H), 5.93 (s, 1H), 4.51-4.48 (t, 1H), 4.03-4.01 (t,2H), 3.75-3.69 (m, 4H), 3.14 (s, 3H), 3.00-2.96 (t, 2H), 2.92-2.85 (m,1H), 1.24-1.22 (d, 6H) ppm; MS (apci, m/z)=390.2 (M+H).

Example 19

8-((2-chloro-4-cyclopropylphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((2-chloro-4-cyclopropylphenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 7 of Method A, substituting2-chloro-4-cyclopropylaniline in place of 2-fluoro-4-(methylthio)anilineto afford2-(2-(tert-butoxy)ethoxy)-8-((2-chloro-4-cyclopropylphenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(21 mg, 41%). MS (apci, m/z)=460.2 (M+H).

Step 2. Preparation of8-((2-chloro-4-cyclopropylphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 8 of Method A, substituting2-(2-(tert-butoxy)ethoxy)-8-((2-chloro-4-cyclopropylphenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionein place of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dioneto afford8-((2-chloro-4-cyclopropylphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(12 mg, 65%). ¹H NMR (400 MHz, CDCl₃) δ 11.13 (s, 1H), 7.14 (d, 1H),6.91-6.88 (dd, 1H), 6.69-6.67 (d, 1H), 5.95 (s, 1H), 4.47-4.44 (t, 1H),4.04-4.01 (t, 2H), 3.75-3.69 (m, 4H), 3.09 (s, 3H), 3.01-2.98 (t, 2H),1.89-1.82 (m, 1H), 1.02-0.97 (m, 2H), 0.69-0.65 (m, 2H) ppm; MS (apci,m/z)=404.1 (M+H).

Example 20

8-((4-acetyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((4-ethynyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.2-Fluoro-4-((trimethylsilyl)ethynyl)aniline (30.2 mg, 0.145 mmol) wasdissolved in THF (1.0 mL, 0.139 mmol) and cooled to −78° C. under N₂atmosphere. LiHMDS (0.277 mL, 1N/THF, 0.277 mmol) was added dropwise,and the mixture was stirred for 45 minutes at the same temperature.Methyl4-(2-((2-(tert-butoxy)ethoxy)amino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(50 mg, 0.139 mmol) was dissolved in THF (1.0 mL, 0.139 mmol) and addeddropwise to the reaction mixture. After stirring for 10 minutes, LiHMDS(0.160 mL, 1N/THF, 0.160 mmol) was added and the reaction was stirredfor 1 hour at the same temperature. The reaction mixture was quenchedwith saturated NH₄Cl (25 mL), then extracted with EtOAc (3×25 mL). Thecombined organic layers were dried over Na₂SO₄, filtered, andconcentrated. The material was dissolved in THF (1.0 mL, 0.139 mmol) andtreated with TBAF (0.1 mL, 1N/THF, 0.100 mmol) at ambient temperaturefor 30 minutes. The mixture was partitioned between EtOAc (25 mL) andsaturated NH₄Cl (25 mL). The organic layer was removed and the aqueouslayer was washed with EtOAc (2×25 mL). The combined organic layers weredried over Na₂SO₄, filtered, and concentrated. The residue was purifiedby column chromatography, eluting with 0-15% (20% MeOH/DCM)/DCM toafford2-(2-(tert-butoxy)ethoxy)-8-((4-ethynyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(19 mg, 32%). MS (apci, m/z)=428.2 (M+H).

Step 2. Preparation of8-((4-acetyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 8 of Method A, substituting2-(2-(tert-butoxy)ethoxy)-8-((4-ethynyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionein place of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dioneto afford8-((4-acetyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(11 mg, 64%). ¹H NMR (400 MHz, CDCl₃) δ 11.19 (s, 1H), 7.76-7.73 (dd,1H), 7.71-7.69 (dd, 1H), 6.87-6.83 (t, 1H), 6.07 (s, 1H), 4.31 (br s,1H), 4.05-4.03 (m, 2H), 3.76-3.73 (m, 4H), 3.24 (s, 3H), 3.03-3.00 (t,2H), 2.58 (s, 3H) ppm; MS (apci, m/z)=390.1 (M+H).

Example 21

8-((2-chloro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((2-chloro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 7 of Method A, substituting2-chloro-4-(methylthio)aniline in place of2-fluoro-4-(methylthio)aniline to afford2-(2-(tert-butoxy)ethoxy)-8-((2-chloro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(15 mg, 29%). MS (apci, m/z)=466.1 (M+H).

Step 2. Preparation of8-((2-chloro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 8 of Method A, substituting2-(2-(tert-butoxy)ethoxy)-8-((2-chloro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionein place of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dioneto afford8-((2-chloro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(9 mg, 68%). ¹H NMR (400 MHz, CDCl₃) δ 11.15 (s, 1H), 7.33 (d, 1H),7.09-7.06 (dd, 1H), 6.71-6.69 (d, 1H), 5.97 (s, 1H), 4.42 (br s, 1H),4.04-4.02 (t, 2H), 3.75-3.71 (m, 4H), 3.12 (s, 3H), 3.01-2.98 (t, 2H),2.48 (s, 3H) ppm; MS (apci, m/z)=410.1 (M+H).

Example 22

8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 7 of Method A, substituting4-(difluoromethoxy)-2-fluoroaniline in place of2-fluoro-4-(methylthio)aniline to afford2-(2-(tert-butoxy)ethoxy)-8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(17 mg, 33%). MS (apci, m/z)=470.2 (M+H).

Step 2.8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 8 of Method A, substituting2-(2-(tert-butoxy)ethoxy)-8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionein place of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dioneto afford8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(10 mg, 67%). ¹H NMR (400 MHz, CDCl₃) δ 11.18 (s, 1H), 7.00-6.91 (m,3H), 6.68-6.31 (t, 1H), 5.98 (s, 1H), 4.43-4.40 (t, 1H), 4.04-4.02 (t,2H), 3.75-3.71 (m, 4H), 3.16 (s, 3H), 3.01-2.98 (t, 2H) ppm; MS (apci,m/z)=414.1 (M+H).

Example 23

8-((2-chloro-4-ethylphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((2-chloro-4-ethylphenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 7 of Method A, substituting2-chloro-4-ethylaniline in place of 2-fluoro-4-(methylthio)aniline toafford2-(2-(tert-butoxy)ethoxy)-8-((2-chloro-4-ethylphenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(9 mg, 14%). MS (apci, m/z)=448.2 (M+H).

Step 2.8-((2-chloro-4-ethylphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 8 of Method A, substituting2-(2-(tert-butoxy)ethoxy)-8-((2-chloro-4-ethylphenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionein place of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dioneto afford8-((2-chloro-4-ethylphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(6 mg, 80%). ¹H NMR (400 MHz, CDCl₃) δ 11.13 (s, 1H), 7.29 (d, 1H),7.02-7.00 (dd, 1H), 6.72-6.70 (d, 1H), 5.95 (s, 1H), 4.46 (br s, 1H),4.04-4.01 (m, 2H), 3.75-3.70 (m, 4H), 3.10 (s, 3H), 3.01-2.97 (t, 2H),2.65-2.59 (q, 2H), 1.24-1.21 (t, 3H) ppm; MS (apci, m/z)=392.1 (M+H).

Example 24

8-((2-fluoro-4-(trifluoromethoxy)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(trifluoromethoxy)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.

Prepared according to Example 6, Step 7 of Method A, substituting2-fluoro-4-(trifluoromethoxy)aniline in place of2-fluoro-4-(methylthio)aniline to afford2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(trifluoromethoxy)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(22 mg, 41%). MS (apci, m/z)=488.2 (M+H).

Step 2.8-((2-fluoro-4-(trifluoromethoxy)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 8 of Method A, substituting2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(trifluoromethoxy)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionein place of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dioneto afford8-((2-fluoro-4-(trifluoromethoxy)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(13 mg, 67%). ¹H NMR (400 MHz, CDCl₃) δ 11.17 (s, 1H), 7.08-7.06 (d,1H), 7.02-6.99 (d, 1H), 6.94-6.90 (t, 1H), 6.01 (s, 1H), 4.40-4.36 (t,1H), 4.05-4.02 (t, 2H), 3.75-3.72 (t, 4H), 3.18 (s, 3H), 3.02-2.99 (t,2H) ppm; MS (apci, m/z)=432.1 (M+H).

Example 25

8-((4-((difluoromethyl)thio)-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((4-((difluoromethyl)thio)-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 7 of Method A, substituting4-((difluoromethyl)thio)-2-fluoroaniline in place of2-fluoro-4-(methylthio)aniline to afford2-(2-(tert-butoxy)ethoxy)-8-((4-((difluoromethyl)thio)-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(19 mg, 35%). δ 11.31 (s, 1H), 7.40-7.37 (dd, 1H), 7.30-7.28 (m, 1H),6.94-6.66 (t, 1H), 6.80-6.76 (t, 1H), 6.04 (s, 1H), 4.17-4.14 (m, 2H),3.82-3.79 (t, 2H), 3.62-3.59 (m, 2H), 3.24 (s, 3H), 3.01-2.97 (t, 2H),1.21 (s, 9H) ppm. MS (apci, m/z)=486.2 (M+H).

Step 2.8-((4-((difluoromethyl)thio)-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, Step 8 of Method A, substituting2-(2-(tert-butoxy)ethoxy)-8-((4-((difluoromethyl)thio)-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionein place of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dioneto afford8-((4-((difluoromethyl)thio)-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(11 mg, 57%). ¹H NMR (400 MHz, CDCl₃) δ 11.16 (s, 1H), 7.42-7.39 (dd,1H), 7.33-7.32 (d, 1H), 6.96-6.68 (t, 1H), 6.87-6.82 (t, 1H), 6.04 (s,1H), 4.37-4.33 (t, 1H), 4.05-4.02 (t, 2H), 3.75-3.72 (t, 4H), 3.22 (s,3H), 3.03-2.99 (t, 2H) ppm; MS (apci, m/z)=430.1 (M+H).

Example 26

8-((4-bromo-2-fluorophenyl)amino)-2-isopropoxy-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 6, steps 1-7 of Method A, substituting0-isopropylhydroxylamine hydrochloride in place ofO-(2-(tert-butoxy)ethyl)hydroxylamine hydrochloride in step 5 and4-bromo-2-fluoroaniline in place of 2-fluoro-4-(methylthio)aniline instep 7 to afford8-((4-bromo-2-fluorophenyl)amino)-2-isopropoxy-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(10.3 mg, 14%). ¹H NMR (400 MHz, CDCl₃) δ 11.37 (s, 1H), 7.30 (dd, 1H),7.21 (dq, 1H), 6.73 (t, 1H), 6.00 (s, 1H), 4.33 (m, 1H), 3.68 (t, 2H),3.18 (s, 3H), 2.99 (t, 2H), 1.30 (d, 6H) ppm; MS (apci, m/z)=424.1,426.1 (M+H).

Example 27

8-((2-fluoro-4-iodophenyl)amino)-2-isopropoxy-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 6, steps 1-7 of Method A, substitutingO-isopropylhydroxylamine hydrochloride in place ofO-(2-(tert-butoxy)ethyl)hydroxylamine hydrochloride in step 5 and2-fluoro-4-iodoaniline in place of 2-fluoro-4-(methylthio)aniline instep 7 to afford8-((2-fluoro-4-iodophenyl)amino)-2-isopropoxy-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(15.2 mg, 23%). ¹H NMR (400 MHz, CDCl₃) δ 11.36 (s, 1H), 7.46 (dd, 1H),7.38 (dt, 1H), 6.58 (t, 1H), 6.00 (s, 1H), 4.33 (m, 1H), 3.68 (t, 2H),3.19 (s, 3H), 2.99 (t, 2H), 1.30 (d, 6H) ppm; MS (apci, m/z)=472.1(M+H).

Example 28

2-ethoxy-8-((2-fluoro-4-iodophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 6, steps 1-7 of Method A, substitutingO-ethylhydroxylamine hydrochloride in place ofO-(2-(tert-butoxy)ethyl)hydroxylamine hydrochloride in step 5 and2-fluoro-4-iodoaniline in place of 2-fluoro-4-(methylthio)aniline instep 7 to afford2-ethoxy-8-((2-fluoro-4-iodophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(24.3 mg, 25%). ¹H NMR 11.33 (s, 1H), 7.46 (d, 1H), 7.38 (d, 1H), 6.57(t, 1H), 6.00 (s, 1H), 4.07 (q, 2H), 3.72 (t, 2H), 3.19 (s, 3H), 2.99(t, 2H), 1.31 (t, 3H) ppm; MS (apci, m/z)=458.0 (M+H).

Example 29

8-((2-fluoro-4-iodophenyl)amino)-2-methoxy-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 6, steps 1-7 of Method A, substitutingO-methylhydroxylamine hydrochloride in place ofO-(2-(tert-butoxy)ethyl)hydroxylamine hydrochloride in step 5 and2-fluoro-4-iodoaniline in place of 2-fluoro-4-(methylthio)aniline instep 7 to afford8-((2-fluoro-4-iodophenyl)amino)-2-methoxy-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(16.7 mg, 33%). ¹H NMR (400 MHz, CDCl₃) δ 11.32 (s, 1H), 7.47 (d, 1H),7.39 (d, 1H), 6.58 (t, 1H), 6.00 (s, 1H), 3.84 (s, 3H), 3.73 (t, 2H),3.19 (s, 3H), 3.00 (t, 2H) ppm; MS (apci, m/z)=444.0 (M+H).

Example 30

2-(tert-butoxy)-8-((2-fluoro-4-iodophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Steps 1-7. Preparation of methyl4-(2-(tert-butoxyamino)ethyl)-2-((2-fluoro-4-iodophenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.Prepared according to Example 6, steps 1-7, substitutingO-(tert-butyl)hydroxylamine hydrochloride in place ofO-(2-(tert-butoxy)ethyl)hydroxylamine hydrochloride in step 5 and2-fluoro-4-iodoaniline in place of 2-fluoro-4-(methylthio)aniline instep 7 to afford methyl4-(2-(tert-butoxyamino)ethyl)-2-((2-fluoro-4-iodophenyl)amino)-1- ofMethod A methyl-6-oxo-1,6-dihydropyridine-3-carboxylate (40.3 mg, 62%).MS (apci, m/z)=518.1 (M+H).

Step 8. Preparation of2-(tert-butoxy)-8-((2-fluoro-4-iodophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.A solution of methyl4-(2-(tert-butoxyamino)ethyl)-2-((2-fluoro-4-iodophenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(40.3 mg, 0.078 mmol) in THF (1 mL) was treated with Et₃N (10.9 μL,0.078 mmol). The reaction mixture was heated to 50° C. for 1 hour. MoreEt₃N (10.9 μL, 0.078 mmol) was added to the reaction and left to stir at50° C. for another hour. An additional aliquot of Et₃N (10.9 μL, 0.078mmol) was added and the reaction was stirred at 50° C. for 5 days. Thereaction mixture was partitioned between water (40 mL) and EtOAc (20mL). The aqueous layer was washed with EtOAc (2×15 mL). The combinedorganic layers were washed with brine (25 mL), dried over sodiumsulfate, filtered, and concentrated. The residue was purified by reversephase HPLC (5-95% acetonitrile/water/0.1% TFA over 20 minutes) and theclean fractions were combined, washed with saturated bicarbonate, andextracted with EtOAc (3×15 mL). The combined organic layers were washedwith saturated bicarbonate (20 mL) and brine (25 mL), dried over sodiumsulfate, filtered, and concentrated in vacuo to afford2-(tert-butoxy)-8-((2-fluoro-4-iodophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(16.5 mg, 43%). ¹H NMR (400 MHz, CDCl₃) δ 11.49 (s, 1H), 7.46 (dd, 1H),7.39 (td, 1H), 6.59 (t, 1H), 5.99 (s, 1H), 3.70-3.60 (m, 2H), 3.18 (s,3H), 3.10-2.87 (m, 2H), 1.34 (s, 9H) ppm; MS (apci, m/z)=486.1 (M+H).

Example 31

(S)-8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxypropoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of methyl(S,E)-2-chloro-4-(2-((2-hydroxypropoxy)imino)ethyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.Methyl(Z)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(200 mg, 0.736 mmol) and(S)-O-(2-((tert-butyldimethylsilyl)oxy)propyl)hydroxylamine (151 mg,0.736) were taken up in 1,4-dioxane (5 mL). TEA (103 μL, 0.736 mmol) andHCl (368 μL, 4N/dioxane, 1.47 mmol) were added. The suspension washeated to 60° C. for 3 hours. The reaction mixture was filtered andconcentrated. The residue was purified by column chromatography elutingwith 0-80% EtOAc/hexanes to afford methyl(S,E)-2-chloro-4-(2-((2-hydroxypropoxy)imino)ethyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(66.6 mg, 29%). MS (apci, m/z)=317.1 (M+H).

Step 2. Preparation of methyl(S,E)-2-chloro-1-methyl-6-oxo-4-(6,8,8,9,9-pentamethyl-4,7-dioxa-3-aza-8-siladec-2-en-1-yl)-1,6-dihydropyridine-3-carboxylate.Methyl(S,E)-2-chloro-4-(2-((2-hydroxypropoxy)imino)ethyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(66.6 mg, 0.210 mmol) and imidazole (0.043 g, 0.631 mmol) were dissolvedin DMF (2 mL) and cooled to 0° C. TBS-CI (63.4 mg, 0.421 mmol) wassubsequently added and the reaction was allowed to warm to roomtemperature over 2 hours. The reaction was diluted with water (40 mL)and EtOAc (20 mL). The aqueous layer was washed with EtOAc (2×15 mL).The combined organic layers were washed with water (5×20 mL) and brine(25 mL), dried over sodium sulfate, filtered and concentrated. Theresidue was purified by column chromatography eluting with 0-40%EtOAc/hexanes to afford methyl(S,E)-2-chloro-1-methyl-6-oxo-4-(6,8,8,9,9-pentamethyl-4,7-dioxa-3-aza-8-siladec-2-en-1-yl)-1,6-dihydropyridine-3-carboxylate(63.4 mg, 70%). MS (apci, m/z)=431.2 (M+H).

Step 3. Preparation of methyl(S)-2-chloro-1-methyl-6-oxo-4-(6,8,8,9,9-pentamethyl-4,7-dioxa-3-aza-8-siladecyl)-1,6-dihydropyridine-3-carboxylate.Methyl(S,E)-2-chloro-1-methyl-6-oxo-4-(6,8,8,9,9-pentamethyl-4,7-dioxa-3-aza-8-siladec-2-en-1-yl)-1,6-dihydropyridine-3-carboxylate(63.4 mg, 0.147 mmol) was dissolved in 2-propanol (1 mL). Sodiumcyanoborohydride (46.2 mg, 0.735 mmol) and acetic acid (42.1 μL, 0.735mmol) were added. The mixture was stirred at room temperature for 16hours. The reaction mixture was partitioned between saturatedbicarbonate (30 mL) and EtOAc (20 mL). The aqueous layer was washed withEtOAc (2×15 mL). The combined organic layers were washed with brine (20mL), dried over sodium sulfate, filtered, and concentrated. The residuewas purified by column chromatography eluting with 0-40% EtOAc/hexanesto afford methyl(S)-2-chloro-1-methyl-6-oxo-4-(6,8,8,9,9-pentamethyl-4,7-dioxa-3-aza-8-siladecyl)-1,6-dihydropyridine-3-carboxylate(30.6 mg, 48%). MS (apci, m/z)=433.2 (M+H).

Step 4. Preparation of(S)-2-(2-((tert-butyldimethylsilyl)oxy)propoxy)-8-((2-fluoro-4-iodophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of 2-fluoro-4-iodoaniline (16.9 mg, 0.071 mmol) inanhydrous THF (1 mL) at −78° C. under N₂ was added LiHMDS (0.141 ml,1M/THF, 0.141 mmol). The mixture was stirred for 45 minutes then asolution of methyl(S)-2-chloro-1-methyl-6-oxo-4-(6,8,8,9,9-pentamethyl-4,7-dioxa-3-aza-8-siladecyl)-1,6-dihydropyridine-3-carboxylate(30.6 mg, 0.071 mmol) in THF (0.5 mL) was added. The mixture was stirredat −78° C. for 10 minutes. The mixture was quenched with saturated NH₄Cl(30 mL), stirred at ambient temperature for 10 minutes, then extractedwith EtOAc (3×10 mL). The combined organic phases were washed with brine(20 mL), dried over sodium sulfate, filtered and concentrated. Theresidue was purified by column chromatography eluting with 0-50%EtOAc/hexanes to afford(S)-2-(2-((tert-butyldimethylsilyl)oxy)propoxy)-8-((2-fluoro-4-iodophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(15.2 mg, 36%). MS (apci, m/z)=602.2 (M+H).

Step 5. Preparation of(S)-8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxypropoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.(S)-2-(2-((tert-butyldimethylsilyl)oxy)propoxy)-8-((2-fluoro-4-iodophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(15.2 mg, 0.025 mmol) was dissolved in MeOH (0.5 mL). The reactionmixture was treated with HCl (12.6 μL, 4N/dioxane, 0.051 mmol) andstirred for 1 hour. The reaction mixture was concentrated and theresidue was purified by reverse phase HPLC (5-95%acetonitrile/water/0.1% TFA over 20 minutes) and the clean fractionswere combined, washed with saturated bicarbonate, and extracted withEtOAc (3×20 mL). The combined organic layers were washed with saturatedbicarbonate (15 mL) and brine (15 mL), dried over sodium sulfate,filtered, and concentrated to afford(S)-8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxypropoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(6.8 mg, 55%). ¹H NMR (400 MHz, CDCl₃) δ 11.16 (s, 1H), 7.48 (dd, 1H),7.42 (dt, 1H), 6.62 (t, 1H), 6.00 (s, 1H), 4.05-3.91 (m, 2H), 3.78-3.60(m, 3H), 3.17 (s, 3H), 3.10-2.90 (m, 2H), 1.15 (d, 3H) ppm; MS (apci,m/z)=488.0 (M+H).

Example 32

(R)-8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxypropoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 31, substituting(R)-O-(2-((tert-butyldimethylsilyl)oxy)propyl)hydroxylamine in place of(S)-O-(2-((tert-butyldimethylsilyl)oxy)propyl)hydroxylamine in step 1 toafford(R)-8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxypropoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(8.6 mg, 49%). ¹H NMR (400 MHz, CDCl₃) δ 11.16 (s, 1H), 7.48 (dd, 1H),7.42 (dt, 1H), 6.62 (t, 1H), 5.99 (s, 1H), 4.05-3.91 (m, 2H), 3.78-3.60(m, 3H), 3.17 (s, 3H), 3.09-2.90 (m, 2H), 1.15 (d, 3H) ppm; MS (apci,m/z)=488.0 (M+H).

Example 33

(S)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxypropoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 31, substituting 4-bromo-2-fluoroanilinein place of 2-fluoro-4-iodoaniline in step 4 to afford(S)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxypropoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(9.2 mg, 55%). ¹H NMR (400 MHz, CDCl₃) δ 11.16 (s, 1H), 7.32 (dd, 1H),7.26-7.22 (m, 1H), 6.78 (t, 1H), 5.99 (s, 1H), 4.05-3.92 (m, 2H),3.78-3.60 (m, 3H), 3.17 (s, 3H), 3.10-2.90 (m, 2H), 1.15 (d, 3H) ppm; MS(apci, m/z)=440.1, 442.1 (M+H).

Example 34

(R)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxypropoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 31, substituting(R)-O-(2-((tert-butyldimethylsilyl)oxy)propyl)hydroxylamine in place of(S)-O-(2-((tert-butyldimethylsilyl)oxy)propyl)hydroxylamine in step 1and 4-bromo-2-fluoroaniline in place of 2-fluoro-4-iodoaniline in step 4to afford(R)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxypropoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(9.2 mg, 55%). ¹H NMR (400 MHz, CDCl₃) δ 11.17 (s, 1H), 7.32 (dd, 1H),7.24 (dt, 1H), 6.78 (t, 1H), 5.99 (s, 1H), 4.06-3.91 (m, 2H), 3.78-3.60(m, 3H), 3.17 (s, 3H), 3.10-2.90 (m, 2H), 1.15 (d, 3H) ppm; MS (apci,m/z)=440.0 (M+H).

Example 35

(R)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxypropoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Steps 1-4: Preparation of(R)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-((tert-butyldimethylsilyl)oxy)propoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 31, steps 1-4 substituting(R)-O-(2-((tert-butyldimethylsilyl)oxy)propyl)hydroxylamine in place of(S)-O-(2-((tert-butyldimethylsilyl)oxy)propyl)hydroxylamine in step 1 toafford(R)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-((tert-butyldimethylsilyl)oxy)propoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(63.5 mg, 62%). MS (apci, m/z)=554.1 (M+H).

Step 5. Preparation of(R)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxypropoxy)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.(R)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-((tert-butyldimethylsilyl)oxy)propoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(42.3 mg, 0.076 mmol) was dissolved in THF (1 mL) and MeOH (1 mL).p-Toluenesulfonic acid monohydrate (21.8 mg, 0.114 mmol) was added.After 5 minutes, N-iodosuccinimide (17.2 mg, 0.076 mmol) was added andleft to stir for 30 minutes. The reaction mixture was partitionedbetween EtOAc (15 mL) and saturated bicarbonate (30 mL) and the aqueouslayer was extracted with EtOAc (2×10 mL). The combined organic phaseswere washed with brine (25 mL), dried over sodium sulfate, filtered, andconcentrated. The residue was purified by column chromatography elutingwith 0-50% EtOAc/heptane to afford(R)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxypropoxy)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(23.8 mg, 54%). MS (apci, m/z)=566.0 (M+H).

Step 6. Preparation of(R)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxypropoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of(R)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxypropoxy)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(21.7 mg, 0.038 mmol) in THF (400 μL) at 0° C. was addedPd(Pt-Bu₃)₂(1.96 mg, 0.004 mmol) followed by methylzinc(II) chloride(19.4 μL, 0.039 mmol). The mixture was removed from the ice bath andstirred for 5 minutes. The mixture was partitioned between saturatedNH₄Cl (20 mL) and EtOAc (15 mL). The aqueous layer was extracted withEtOAc (2×10 mL). The combined organic phases were washed with brine (20mL), dried over sodium sulfate, filtered, and concentrated. The residuewas purified by reverse phase HPLC (5-95% acetonitrile/water/0.1% TFAover 20 minutes) and the clean fractions were combined, washed withsaturated bicarbonate, and extracted with EtOAc (3×10 mL). The combinedorganic layers were washed with saturated bicarbonate (25 mL) and brine(30 mL), dried over sodium sulfate, filtered, and concentrated to afford(R)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxypropoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(7.5 mg, 43%). ¹H NMR (400 MHz, CDCl₃) δ 10.97 (s, 1H), 7.30 (dd, 1H),7.19 (dt, 1H), 6.68 (t, 1H), 4.05-3.91 (m, 2H), 3.71 (m, 2H), 3.64 (m,1H), 3.22 (s, 3H), 3.13-3.04 (m, 1H), 3.03-2.92 (m, 1H), 2.08 (s, 3H),1.14 (d, 3H) ppm; MS (apci, m/z)=454.1 (M+H).

Example 36

8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of 4-bromo-2-fluoroaniline (88 mg, 0.47 mmol) in anhydrousTHF (5 mL) at −78° C. under N₂ was added LiHMDS (694 mL, 1.0 M/THF,0.694 mmol). The mixture was stirred for 45 min, then a solution ofmethyl4-(2-((2-(tert-butoxy)ethoxy)amino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(167 mg, 0.46 mmol) in THF (2 mL) was added. Stirring was continued at−78° C. for 10 minutes, then LiHMDS (300 mL, 1.0 M/THF, 0.300 mmol) wasadded. After stirring for another 10 minutes at −78° C., LiHMDS (100 mL,1.0 M/THF, 0.100 mmol) was added, and stirring continued for 10 minutes.The mixture was quenched with saturated NH₄Cl (10 mL), stirred atambient temperature for 10 minutes, then extracted with EtOAc (3×10 mL).The combined organic phases were washed with brine (10 mL), dried overNa₂SO₄, filtered and concentrated. The residue was purified by columnchromatography, eluting with 0-15% (20% MeOH/DCM)/DCM, to afford8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(78 mg, 35%). MS (apci, m/z)=482.1 (M+H).

Step 2. Preparation of8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(78 mg, 0.16 mmol) in THF (2 mL) and MeOH (2 mL) was addedp-toluenesulfonic acid monohydrate (46 mg, 0.24 mmol). After 5 minutes,N-iodosuccinimide (36 mg, 0.16 mmol) was added. After stirring atambient temperature for 30 minutes, the reaction mixture was partitionedbetween EtOAc (10 mL) and saturated NaHCO₃ (10 mL). The aqueous wasextracted with EtOAc (2×10 mL) and the combined organic phases werewashed with brine (10 mL), dried over Na₂SO₄, filtered, andconcentrated. The residue was purified by column chromatography elutingwith 0-30% EtOAc/hexanes to afford8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(62 mg, 63%). ¹H NMR (400 MHz, CDCl₃) δ 11.54 (s, 1H), 7.31 (dd, 1H),7.22 (m, 1H), 6.73 (t, 1H), 4.17-4.13 (m, 2H), 3.80 (t, 2H), 3.63-3.59(m, 2H), 3.27 (s, 3H), 3.17 (t, 2H), 1.22 (s, 9H) ppm; MS (apci,m/z)=608.0 (M+H).

Step 3. Preparation of8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(62 mg, 0.10 mmol) in anhydrous THF (2 mL) at 0° C. was addedPd(Pt-Bu₃)₂(5 mg, 0.01 mmol) followed by methylzinc(II)chloride (61 mL,2M/THF, 0.12 mmol). The mixture was removed from the ice bath andstirred for 5 minutes, then partitioned between saturated NH₄Cl (10 mL)and EtOAc (10 mL). The aqueous layer was extracted with EtOAc (2×10 mL)and the combined organic phases were washed with brine (10 mL), driedover Na₂SO₄, filtered and concentrated. The residue was purified bycolumn chromatography eluting with 0-60% EtOAc/hexanes to afford8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(39 mg, 77%). ¹H NMR (400 MHz, CDCl₃) δ 11.11 (s, 1H), 7.28 (dd, 1H),7.16 (dt, 1H), 6.61 (t, 1H), 4.17-4.13 (m, 2H), 3.78 (t, 2H), 3.64-3.59(m, 2H), 3.25 (s, 3H), 3.02 (t, 2H), 2.09 (s, 3H), 1.22 (s, 9H) ppm; MS(apci, m/z)=496.1 (M+H).

Step 4. Preparation of8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(39 mg, 0.079 mmol) in acetonitrile (0.5 mL) was added phosphoric acid(0.5 mL). The mixture was warmed to 60° C. for 1 hour then cooled toambient temperature. The mixture was stirred with saturated NaHCO₃ (10mL) and EtOAc (10 mL) for 10 minutes and the layers separated. Theaqueous layer was extracted with EtOAc (2×10 mL) and the combinedorganic phases were washed with brine (10 mL), dried over Na₂SO₄,filtered and concentrated. The residue was purified by reverse phaseHPLC (0-95% acetonitrile/water/0.1% TFA over 20 min) and clean fractionswere worked up with saturated NaHCO₃/EtOAc to afford8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(18 mg, 52%). ¹H NMR (400 MHz, CDCl₃) δ 10.96 (s, 1H), 7.31 (dd, 1H),7.20 (dt, 1H), 6.68 (t, 1H), 4.05-4.01 (m, 2H), 3.75-3.69 (m, 4H), 3.23(s, 3H), 3.03 (t, 2H), 2.08 (s, 3H) ppm; MS (apci, m/z)=440.0 (M+H).

Example 37

8-((4-bromo-2-fluorophenyl)amino)-5-chloro-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 6, steps 1-7 of Method A, substituting4-bromo-2-fluoroaniline in place of 2-fluoro-4-(methylthio)aniline instep 7 to afford8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(28 mg, 35%). MS (apci, m/z)=482.1 (M+H).

Step 2. Preparation of8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-5-chloro-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(28 mg, 0.058 mmol) in DMF (0.5 mL) was added NCS (8 mg, 0.058 mmol).The mixture was stirred at ambient temperature for 1 hour, then at 50°C. for 2 hours. The cooled mixture was partitioned between saturatedNaHCO₃ (10 mL) and EtOAc (10 mL) and the aqueous layer was extractedwith EtOAc (2×10 mL). The combined organic phases were washed with water(5×10 mL) and brine (10 mL) then dried over Na₂SO₄, filtered andconcentrated. The residue was purified by column chromatography elutingwith 0-50% EtOAc/hexanes to afford8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-5-chloro-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(14 mg, 47%). MS (apci, m/z)=516.0 (M+H).

Step 3. Preparation of8-((4-bromo-2-fluorophenyl)amino)-5-chloro-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-5-chloro-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(14 mg, 0.027 mmol) in acetonitrile (0.5 mL) was added phosphoric acid(0.5 mL). The mixture was warmed to 60° C. for 30 minutes then cooled toambient temperature. The mixture was stirred with saturated NaHCO₃ (10mL) and EtOAc (10 mL) for 10 minutes and the layers separated. Theaqueous layer was extracted with EtOAc (2×10 mL) and the combinedorganic phases were washed with brine (10 mL), dried over Na₂SO₄,filtered and concentrated. The residue was purified by columnchromatography eluting with 0-15% (20% MeOH/DCM)/DCM to afford8-((4-bromo-2-fluorophenyl)amino)-5-chloro-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(11 mg, 88%). ¹H NMR (400 MHz, CDCl₃) δ 11.22 (s, 1H), 7.33 (dd, 1H),7.26 (dt, 1H), 6.78 (t, 1H), 4.07-4.02 (m, 2H), 3.78-3.70 (m, 4H), 3.25(s, 3H), 3.22 (t, 2H) ppm; MS (apci, m/z)=460.0 (M+H).

Example 38

5-chloro-8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 37, substituting 4-iodo-2-fluoroaniline inplace of 4-bromo-2-fluoroaniline in Step 1 to afford5-chloro-8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(10 mg, 93%). ¹H NMR (400 MHz, CDCl₃) δ 11.21 (s, 1H), 7.49 (dd, 1H),7.43 (dt, 1H), 6.63 (t, 1H), 4.25-4.17 (m, 1H), 4.07-4.02 (m, 2H),3.79-3.70 (m, 4H), 3.25 (s, 3H), 3.22 (t, 2H) ppm; MS (apci, m/z)=508.0(M+H).

Example 39

8-((4-bromo-2-fluorophenyl)amino)-5-fluoro-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-5-fluoro-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.A solution of8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(77 mg, 0.16 mmol) and1-(chloromethyl)-4-fluoro-1,4-diazabicyclo[2.2.2]octane-1,4-diiumtetrafluoroborate (57 mg, 0.16 mmol) in acetonitrile (9.0 mL) wasstirred at 0° C. for 1 hour. The reaction mixture was diluted with EtOAc(50 mL) and washed with water (25 mL) followed by brine (25 mL). Theorganic phase was dried over Na₂SO₄, filtered and concentrated. Theresidue was purified by reverse phase HPLC (5-95%acetonitrile/water/0.1% TFA over 20 minutes). The fractions containingthe desired product were combined and diluted with saturated NaHCO₃ (15mL) then extracted with EtOAc (2×25 mL). The organic phases werecombined and dried over Na₂SO₄, filtered and concentrated to afford8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-5-fluoro-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(10 mg, 13%). MS (apci, m/z)=500.1 (M+H).

Step 2. Preparation of8-((4-bromo-2-fluorophenyl)amino)-5-fluoro-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.A solution of8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-5-fluoro-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(13 mg, 0.026 mmol) and conc. H₃PO₄ (0.20 mL, 3.0 mmol) in acetonitrile(1.0 mL) was stirred at 60° C. for 1 hour. The reaction mixture wasconcentrated then diluted with K₃PO₄ (1M in water, 3.0 mL, 3.0 mmol).The aqueous phase was extracted with EtOAc (2×15 mL) and the organicphases were combined, dried over Na₂SO₄, filtered and concentrated. Theresidue was purified by HPLC (10-95% acetonitrile/water/0.1% TFA). Thefractions containing the desired product were combined and diluted withsaturated NaHCO₃ (15 mL). The aqueous was extracted with EtOAc (2×15 mL)and the organic phases were combined, dried over Na₂SO₄, filtered andconcentrated to afford8-((4-bromo-2-fluorophenyl)amino)-5-fluoro-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(5.0 mg, 43%). 1H NMR (400 MHz, CDCl3) δ 10.79 (s, 1H), 7.34-7.30 (m,1H), 7.25-7.21 (m, 1H), 6.72 (t, 1H), 4.23 (t, 1H), 4.06-4.02 (m, 2H),3.80-3.70 (m, 4H), 3.25 (s, 3H), 3.17-3.12 (m, 2H) ppm; MS (apci,m/z)=444.0 (M+H).

Example 40

8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-iodophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.8-((4-Bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(Prepared according to Example 36, Steps 1-3; 65 mg, 0.13 mmol),copper(I) iodide (6.2 mg, 0.033 mmol), sodium iodide (39 mg, 0.26 mmol),and (1S,2S)-N1,N2-dimethylcyclohexane-1,2-diamine (9.3 mg, 0.065 mmol)were combined in 1,4-dioxane (1 mL) and sparged with argon. The mixturewas heated at 110° C. for 18 hours then at ambient temperature for 48hours. The mixture was partitioned between EtOAc (20 mL) and ammoniumhydroxide (10 mL/water 10 mL). The organic phase was separated and theaqueous extracted with EtOAc (2×10 mL). Combined organic phases werewashed with brine (10 mL), dried over Na₂SO₄, filtered and concentrated.The residue was purified by column chromatography eluting with 0-80%EtOAc/heptane to afford2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-iodophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(49 mg, 69%). MS (apci, m/z)=544.1 (M+H).

Step 2. Preparation of8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-iodophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(49 mg, 0.09 mmol) in acetonitrile (1 mL) was added phosphoric acid (0.5mL). The mixture was warmed to 60° C. for 1 hour then cooled to ambienttemperature. The mixture was stirred with saturated NaHCO₃ (10 mL) andEtOAc (10 mL) for 10 minutes and the layers separated. The aqueous layerwas extracted with EtOAc (2×10 mL) and the combined organic phases werewashed with brine (10 mL), dried over Na₂SO₄, filtered and concentrated.The residue was purified by reverse phase HPLC (5-95%acetonitrile/H₂O/0.1% TFA over 20 min) and clean fractions were workedup with EtOAc/saturated NaHCO₃ to afford8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(23 mg, 52%). ¹H NMR (400 MHz, CDCl₃) δ 10.95 (s, 1H), 7.47 (dd, 1H),7.37 (dt, 1H), 6.52 (t, 1H), 4.05-4.02 (m, 2H), 3.75-3.65 (m, 4H), 3.24(s, 3H), 3.03 (t, 2H), 3.02 (s, 3H) ppm; MS (apci, m/z)=488.0 (M+H).

Example 41

8-((2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of methyl4-bromo-2-((2-fluorophenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.To a solution of 2-fluoroaniline (100 mg, 0.90 mmol) in anhydrous THF (5mL) at −78° C. under N₂ was added LiHMDS (1.34 mL, 1.0 M/THF, 1.34mmol). The mixture was stirred for 45 minutes, then a solution of methyl4-bromo-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate (250mg, 0.89 mmol) in THF (5 mL) was added. The mixture was stirred at −78°C. for 50 minutes. Additional LiHMDS (0.5 mL, 1.0 M/THF, 0.5 mmol) wasadded and stirring continued for another 10 minutes. The mixture wasquenched with saturated NH₄Cl (20 mL), stirred at ambient temperaturefor 10 minutes, then extracted with EtOAc (3×20 mL). The combinedorganic phases were washed with brine (10 mL), dried over sodiumsulfate, filtered, and concentrated to afford methyl4-bromo-2-((2-fluorophenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(291 mg, 92%). MS (apci, m/z)=355.0 (M+H).

Step 2. Preparation of methyl(E)-4-(2-ethoxyvinyl)-2-((2-fluorophenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.Methyl4-bromo-2-((2-fluorophenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(188 mg, 0.529 mmol),methanesulfonato(2-dicyclohexylphosphino-2′,6′-di-i-′propoxy-1,1-biphenyl)′2-amino-1,1-biphenyl-2-yl)palladium(II)(44.3 g, 0.053 mmol), and(E)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (123 μL,0.582 mmol) were suspended in 1,4-dioxane (5 mL) and potassium carbonate(0.397 mL, 2N aq, 0.794 mmol) was added. After degassing with argon, themixture was stirred at ambient temperature for 23 hours. The reactionwas partitioned between water (40 mL) and EtOAc (30 mL). The aqueouslayer was washed with EtOAc (2×15 mL). The combined organic layers werewashed with brine (40 mL), dried over sodium sulfate, and concentrated.The residue was purified by column chromatography eluting with 0-50%EtOAc/hexanes to methyl(E)-4-(2-ethoxyvinyl)-2-((2-fluorophenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(91.5 mg, 50%). MS (apci, m/z)=347.1 (M+H).

Step 3. Preparation of methyl(E)-4-(2-((2-(tert-butoxy)ethoxy)imino)ethyl)-2-((2-fluorophenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.Methyl(E)-4-(2-ethoxyvinyl)-2-((2-fluorophenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(91.5 mg, 0.264 mmol) and O-(2-(tert-butoxy)ethyl)hydroxylaminehydrochloride (44.8 mg, 0.264 mmol) were taken up in 1,4-dioxane (2 mL).TEA (36.8 μL, 0.264 mmol) and HCl (66 μL, 4N/dioxane, 0.264 mmol) wereadded. The suspension was heated to 60° C. After 2 hours, additional HCl(66 μL, 4N/dioxane, 0.264 mmol) was added and stirring continued at 60°C. for another 2 hours. The reaction mixture was filtered andconcentrated. The residue was purified by column chromatography elutingwith 0-40% EtOAc/hexanes to afford methyl(E)-4-(2-((2-(tert-butoxy)ethoxy)imino)ethyl)-2-((2-fluorophenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(67.6 mg, 59%). MS (apci, m/z)=434.2 (M+H).

Step 4. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Methyl(E)-4-(2-((2-(tert-butoxy)ethoxy)imino)ethyl)-2-((2-fluorophenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(67.6 mg, 0.156 mmol) was dissolved in MeOH (1.0 mL) and 2-propanol (1.0mL). Sodium cyanoborohydride (49 mg, 0.780 mmol) and acetic acid (44.6μL, 0.780 mmol) were added, and the mixture was stirred at ambienttemperature for 16 hours. The reaction was heated at 40° C. for 16hours. The reaction was partitioned between saturated bicarbonate (20mL) and EtOAc (10 mL). The aqueous layer was washed EtOAc (2×10 mL). Thecombined organic layers were dried over sodium sulfate, filtered, andconcentrated. The residue was purified by column chromatography elutingwith 0-80% EtOAc/hexanes to afford2-(2-(tert-butoxy)ethoxy)-8-((2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(37.8 mg, 60%). MS (apci, m/z)=404.2 (M+H).

Step 5. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((2-fluorophenyl)amino)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.2-(2-(tert-butoxy)ethoxy)-8-((2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(19 mg, 0.047 mmol) was dissolved in THF (0.5 mL) and MeOH (0.5 mL).p-Toluenesulfonic acid monohydrate (13 mg, 0.071 mmol) was added. After5 minutes, N-Iodosuccinimide (11 mg, 0.047 mmol) was added and left tostir for 1 hour. The reaction mixture was partitioned between EtOAc (20mL) and saturated bicarbonate (40 mL). The aqueous layer was washed withEtOAc (2×15 mL). The combined organic layers were washed with brine (20mL), dried over sodium sulfate, filtered, and concentrated. The residuewas purified by column chromatography eluting with 0-30% EtOAc/hexanesto afford2-(2-(tert-butoxy)ethoxy)-8-((2-fluorophenyl)amino)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(9.9 mg, 40%). MS (apci, m/z)=530.1 (M+H).

Step 6. Preparation of8-((2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.2-(2-(tert-butoxy)ethoxy)-8-((2-fluorophenyl)amino)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(9.9 mg, 0.019 mmol) was dissolved in acetonitrile (0.5 mL) andphosphoric acid (0.5 mL) was added. The solution was heated to 60° C.for 1 hour. The reaction mixture was partitioned between saturatedbicarbonate (15 mL) and EtOAc (10 mL). The aqueous layer was washed withEtOAc (2×8 mL). The combined organic layers were washed with brine (15mL), dried over sodium sulfate, filtered, and concentrated. The residuewas purified by reverse phase HPLC (5-95% acetonitrile/water/0.1% TFAover 20 minutes) and the clean fractions were combined, washed withsaturated bicarbonate, and extracted with EtOAc (3×10 mL). The combinedorganic layers were washed with saturated bicarbonate (20 mL) and brine(20 mL), dried over sodium sulfate, filtered, and concentrated to afford(8-((2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(4.5 mg, 51%). ¹H NMR (400 MHz, CDCl₃) δ 11.41 (s, 1H), 7.21-7.08 (m,3H), 6.94 (t, 1H), 4.04 (t, 2H), 3.76-3.69 (m, 4H), 3.25 (s, 3H), 3.19(t, 2H) ppm; MS (apci, m/z)=474.0 (M+H).

Example 42

5-bromo-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 41, substituting 4-bromo-2-fluoro anilinein place of 2-fluoroaniline in step 1 and N-bromosuccinimide in step 5to afford5-bromo-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(6.5 mg, 29%). ¹H NMR (400 MHz, CDCl₃) δ 11.28 (s, 1H), 7.33 (dd, 1H),7.25 (m, 1H), 6.80 (t, 1H), 4.05 (t, 2H), 3.78-3.70 (m, 4H), 3.25 (s,3H), 3.22 (t, 2H) ppm; MS (apci, m/z)=504.0 (M+H).

Example 43

4-bromo-8-((2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of2-(2-(tert-butoxy)ethoxy)-8-((2-fluorophenyl)amino)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(75 mg, 0.14 mmol) in anhydrous THF (5 mL) at ambient temperature wasadded methylzinc(II)chloride (140 mL, 2M, 0.14 mmol). After stirring for5 minutes, the mixture was concentrated. The residue was purified bycolumn chromatography eluting with 0-80% EtOAc/hexanes to afford2-(2-(tert-butoxy)ethoxy)-8-((2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(38 mg, 64%). MS (apci, m/z)=418.2 (M+H).

Step 2. Preparation of4-bromo-2-(2-(tert-butoxy)ethoxy)-8-((2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of2-(2-(tert-butoxy)ethoxy)-8-((2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(38 mg, 0.09 mmol) in anhydrous DMF (1 mL) was added NBS (17 mg, 0.09mmol). The mixture was stirred at ambient temperature for 2 hours thenpartitioned between saturated NaHCO₃ (10 mL) and EtOAc (10 mL). Theaqueous layer was extracted with EtOAc (2×10 mL) and the combinedorganic phases were washed with water (5×10 mL) and brine (10 mL) thendried over Na₂SO₄, filtered and concentrated. The residue was purifiedby column chromatography eluting with 0-60% EtOAc/hexanes to afford4-bromo-2-(2-(tert-butoxy)ethoxy)-8-((2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(30 mg, 66%). MS (apci, m/z)=496.2 (M+H).

Step 3. Preparation of4-bromo-8-((2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of4-bromo-2-(2-(tert-butoxy)ethoxy)-8-((2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(30 mg, 0.060 mmol) in acetonitrile (0.5 mL) was added phosphoric acid(0.5 mL). The mixture was warmed to 60° C. for 1 hour then cooled andstirred with saturated NaHCO₃ (10 mL) and EtOAc (10 mL) for 10 minutes.Layers were separated and the aqueous layer was extracted with EtOAc(2×10 mL). The combined organic phases were washed with brine (10 mL),dried over Na₂SO₄, filtered and concentrated. The residue was purifiedby column chromatography eluting with 0-15% (20% MeOH/DCM)/DCM to afford4-bromo-8-((2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(10 mg, 38%). ¹H NMR (400 MHz, CDCl₃) δ 11.08 (s, 1H), 7.15-7.05 (m,2H), 6.86 (t, 1H), 5.36 (t, 1H), 4.17-4.04 (m, 3H), 3.99-3.92 (m, 1H),3.88 (dd, 1H), 3.68 (dt, 1H), 3.21 (s, 3H), 2.17 (s, 3H), 1.26 (t, 1H)ppm; MS (apci, m/z)=440.1 (M+H).

Example 44

8-((4-ethyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of methyl(Z)-4-(2-ethoxyvinyl)-2-((4-ethyl-2-fluorophenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.To a solution of 4-ethyl-2-fluoroaniline (155 mg, 1.12 mmol) inanhydrous THF (2 mL) at −78° C. under N₂ was added LiHMDS (2.21 mL,1M/THF, 2.21 mmol). The mixture was stirred for 40 minutes, then asolution of methyl(Z)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(300 mg, 1.10 mmol) in THF (1 mL) was added via syringe. The mixture wasstirred at −78° C. for 45 minutes. The mixture was quenched withsaturated NH₄Cl (35 mL), stirred at ambient temperature for 5 minutes,then extracted with EtOAc (3×15 mL). The combined organic phases werewashed with brine (30 mL), dried over sodium sulfate, filtered, andconcentrated. The residue was purified by column chromatography elutingwith 0-50% EtOAc/heptane to afford methyl(Z)-4-(2-ethoxyvinyl)-2-((4-ethyl-2-fluorophenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(298 mg, 72%). MS (apci, m/z)=375.2 (M+H).

Step 2. Preparation of methyl2-((4-ethyl-2-fluorophenyl)amino)-1-methyl-6-oxo-4-(2-oxoethyl)-1,6-dihydropyridine-3-carboxylate.Methyl(Z)-4-(2-ethoxyvinyl)-2-((4-ethyl-2-fluorophenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(298 mg, 0.797 mmol) was dissolved in DCM (800 μL) and TFA (921 μL, 11.9mmol) was added. The reaction was stirred at ambient temperature for 16hours. The mixture was partitioned between DCM (20 mL) and saturatedsodium bicarbonate (35 mL). The aqueous layer was washed with DCM (2×15mL). The combined organic layers were washed with saturated sodiumbicarbonate (30 mL), dried over sodium sulfate, filtered andconcentrated to afford methyl2-((4-ethyl-2-fluorophenyl)amino)-1-methyl-6-oxo-4-(2-oxoethyl)-1,6-dihydropyridine-3-carboxylate(276 mg, 100%). MS (apci, m/z)=347.1 (M+H).

Step 3. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((4-ethyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Methyl2-((4-ethyl-2-fluorophenyl)amino)-1-methyl-6-oxo-4-(2-oxoethyl)-1,6-dihydropyridine-3-carboxylate(276 mg, 0.797 mmol), O-(2-(tert-butoxy)ethyl)hydroxylaminehydrochloride (117 mg, 0.876 mmol), sodium triacetoxyhydroborate (203mg, 0.956 mmol), and acetic acid (45.6 μL, 0.797 mmol) were taken up inDCE (8 mL). The reaction was stirred at ambient temperature for 1 hour.The reaction was heated to 55° C. for 20 minutes. More sodiumtriacetoxyhydroborate (400 mg) and acetic acid (90 μL) were added to thereaction and stirred at ambient temperature for 16 hours. Sodiumcyanoborohydride (250 mg, 3.98 mmol) and acetic acid (250 μL) were addedto the reaction and allowed to stir at ambient temperature for 3 hours.The solution was partitioned between DCM (40 mL) and saturated sodiumbicarbonate (70 mL). The aqueous layer was washed with DCM (2×20 mL).The combined organic layers were washed with brine (50 mL), dried oversodium sulfate, filtered and concentrated. The residue was purified bycolumn chromatography eluting with 0-100% EtOAc/heptane to afford2-(2-(tert-butoxy)ethoxy)-8-((4-ethyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(137 mg, 40%). MS (apci, m/z)=432.2 (M+H).

Step 4. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((4-ethyl-2-fluorophenyl)amino)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.2-(2-(tert-Butoxy)ethoxy)-8-((4-ethyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(137 mg, 0.317 mmol) was dissolved in THF (1 mL)/MeOH (1 mL).p-Toluenesulfonic acid monohydrate (90.4 mg, 0.475 mmol) was added.After 5 minutes, N-iodosuccinimide (71.3 mg, 0.317 mmol) was added andleft to stir for 30 minutes. The reaction mixture was partitionedbetween EtOAc (20 mL) and saturated sodium bicarbonate (30 mL) and theaqueous layer was extracted with EtOAc (2×10 mL). The combined organicphases were washed with brine (25 mL), dried over sodium sulfate,filtered, and concentrated to afford2-(2-(tert-butoxy)ethoxy)-8-((4-ethyl-2-fluorophenyl)amino)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(143 mg, 81%). MS (apci, m/z)=558.1 (M+H).

Step 5. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((4-ethyl-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of2-(2-(tert-butoxy)ethoxy)-8-((4-ethyl-2-fluorophenyl)amino)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(143 mg, 0.256 mmol) in THF (2.5 mL, 0.256 mmol) at 0° C. was addedPd(Pt-Bu₃)₂ (13.1 mg, 0.026 mmol) followed by methylzinc(II) chloride(130 μL, 0.259 mmol). The mixture was removed from the ice bath andstirred for 45 minutes. The mixture was partitioned between saturatedNH₄Cl (50 mL) and EtOAc (20 mL) and the aqueous layer was extracted withEtOAc (2×15 mL). The combined organic phases were washed with brine (40mL), dried over sodium sulfate, filtered, and concentrated. The residuewas purified by column chromatography eluting with 0-50% EtOAc/heptaneto afford2-(2-(tert-butoxy)ethoxy)-8-((4-ethyl-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(53.7 mg, 47%). MS (apci, m/z)=446.2 (M+H).

Step 6. Preparation of8-((4-ethyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.2-(2-(tert-Butoxy)ethoxy)-8-((4-ethyl-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(53.7 mg, 0.121 mmol) was dissolved in acetonitrile (0.75 mL) andphosphoric acid (0.75 mL) was added. The reaction was heated to 60° C.for 60 minutes. The reaction mixture was partitioned between saturatedsodium bicarbonate (40 mL) and EtOAc (20 mL). The aqueous layer waswashed with EtOAc (2×10 mL). The combined organic layers were washedwith saturated sodium bicarbonate (30 mL) and brine (25 mL), dried oversodium sulfate, filtered, and concentrated. The residue was purified bycolumn chromatography eluting with 0-100% EtOAc/heptane to afford8-((4-ethyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(40.6 mg, 87%). ¹H NMR (400 MHz, CDCl₃) δ 11.01 (s, 1H), 6.95 (dd, 1H),6.89 (dd, 1H), 6.77 (t, 1H), 4.53 (t, 1H), 4.02 (t, 2H), 3.75-3.67 (m,4H), 3.20 (s, 3H), 3.02 (t, 2H), 2.62 (q, 2H), 2.08 (s, 3H), 1.22 (t,3H) ppm; MS (apci, m/z)=390.2 (M+H).

Example 45

8-((4-cyclopropyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 44 substituting4-cyclopropyl-2-fluoroaniline in place of 4-ethyl-2-fluoroaniline instep 1 to afford8-((4-cyclopropyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(30.7 mg, 75%). ¹H NMR (400 MHz, CDCl₃) δ 11.00 (s, 1H), 6.81-6.71 (m,3H), 4.52 (t, 1H), 4.02 (m, 2H), 3.75-3.66 (m, 4H), 3.19 (s, 3H), 3.02(t, 2H), 2.07 (s, 3H), 1.86 (m, 1H), 1.02-0.95 (m, 2H), 0.69-0.63 (m,2H) ppm; MS (apci, m/z)=402.2 (M+H).

Example 46

8-((4-bromo-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionetrifluoroacetate

Step 1. Preparation of methyl(E)-2-((4-bromo-2-fluorophenyl)amino)-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.4-Bromo-2-fluoroaniline (492 mg, 2.59 mmol) and methyl(E)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(670 mg, 2.47 mmol) was dissolved in THF (12.3 mL, 0.2 M) and cooled to−78° C. under a nitrogen atmosphere. LiHMDS (4.9 mL, 1.0 M/THF, 4.9mmol) was added dropwise, then the mixture allowed to warm from −78° C.to ambient temperature over 1 hour. The reaction mixture was quenchedwith saturated aq. NH₄Cl (15 mL), then extracted with EtOAc (2×10 mL).The combined organic phases were dried over MgSO₄, filtered, andconcentrated. The residue was purified by column chromatography elutingwith 0-100% EtOAc/Heptane to afford methyl(E)-2-((4-bromo-2-fluorophenyl)amino)-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(810 mg, 77%). MS (apci, m/z)=425.0 (M+H).

Step 2. Preparation of methyl2-((4-bromo-2-fluorophenyl)amino)-1-methyl-6-oxo-4-(2-oxoethyl)-1,6-dihydropyridine-3-carboxylate.To a solution of methyl(E)-2-((4-bromo-2-fluorophenyl)amino)-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(810 mg, 1.90 mmol) in DCM (5.0 mL) was added TFA (5.0 mL). The solutionwas allowed to stir at ambient temperature for 16 hours, thenconcentrated. The crude mixture was then diluted with DCM (10 mL) andwashed with saturated aq. NaHCO₃ (10 mL). The organic phase was driedover MgSO₄, filtered, and concentrated to afford methyl2-((4-bromo-2-fluorophenyl)amino)-1-methyl-6-oxo-4-(2-oxoethyl)-1,6-dihydropyridine-3-carboxylatewhich was used without further purification (751 mg, 99%). MS (apci,m/z)=397.0, 399.0 (M+H).

Step 3. Preparation of8-((4-bromo-2-fluorophenyl)amino)-2-(4-methoxybenzyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Methyl2-((4-bromo-2-fluorophenyl)amino)-1-methyl-6-oxo-4-(2-oxoethyl)-1,6-dihydropyridine-3-carboxylate(751 mg, 1.89 mmol), (4-methoxyphenyl)methanamine (272 mg, 1.99 mmol),sodium triacetoxyborohydride (441 mg, 2.08 mmol), and acetic acid (11.4mg, 0.189 mmol) were dissolved in DCE (19 mL) and allowed to stir atambient temperature for 1 hour. The reaction was warmed to 60° C. andallowed to stir for 1 hour. The reaction was then cooled to ambienttemperature, diluted with DCM (10 mL) and washed with saturated aq.NaHCO₃ (15 mL). The organic phase was dried over MgSO₄, filtered, andconcentrated. The residue was purified by column chromatography elutingwith 0-100% EtOAc/Heptane to afford8-((4-bromo-2-fluorophenyl)amino)-2-(4-methoxybenzyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(630 mg, 69%). MS (apci, m/z)=486.0 (M+H).

Step 4. Preparation of8-((4-bromo-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionetrifluoroacetate.8-((4-bromo-2-fluorophenyl)amino)-2-(4-methoxybenzyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(630 mg, 1.30 mmol) was dissolved in TFA (8.0 mL) and heated to 80° C.for 16 hours. The reaction was allowed to cool to ambient temperatureand concentrated. The residue was purified by reverse phase HPLC (5-95%acetonitrile/H₂O/0.1% TFA). The fractions containing the clean desiredproduct were lyophilized to afford8-((4-bromo-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionetrifluoroacetate (226 mg, 48%). ¹H NMR (400 MHz, CDCl₃) δ 11.28 (s, 1H),7.32-7.29 (m, 1H), 7.23-7.20 (m, 1H), 6.74 (t, 1H), 6.05 (s, 1H), 5.62(s, 1H), 3.51-3.47 (m, 2H), 3.20 (s, 3H), 2.84 (t, 2H) ppm. MS (apci,m/z)=366.0, 368.0 (M+H).

Example 47

8-((4-bromo-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of8-((4-bromo-2-fluorophenyl)amino)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.8-((4-bromo-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(Prepared according to Example 46; 41 mg, 0.11 mmol) andp-toluenesulfonic acid (29 mg, 0.17 mmol) were dissolved in 1 mL of a1:1 THF/MeOH solution. After 5 minutes, NIS (25 mg, 0.11 mmol) wasadded. The reaction was allowed to stir at ambient temperature for 30minutes. The crude reaction mixture was then partitioned between EtOAc(10 mL) and saturated aq. NaHCO₃ (15 mL) and extracted with EtOAc (2×5mL). The combined organic phases were dried over MgSO₄, filtered, andconcentrated to afford8-((4-bromo-2-fluorophenyl)amino)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionewhich was used without further purification (55 mg, 100%). MS (apci,m/z)=493.9 (M+H).

Step 2. Preparation of8-((4-bromo-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.8-((4-bromo-2-fluorophenyl)amino)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(55 mg, 0.112 mmol) and bis(tri-t-butylphosphine)palladium(0) (5.71 mg,0.0112 mmol) were dissolved in THF (1.1 mL) and methylzinc(II) chloride(55.9 μL, 2M/THF, 0.112 mmol) was added dropwise. The reaction wasallowed to stir at ambient temperature for 1 hour. The reaction mixturewas quenched with saturated aq. NaHCO₃ (2 mL), then extracted with EtOAc(2×5 mL). The combined organic phases were dried over MgSO₄, filtered,and concentrated. The residue was purified by reverse phasechromatography eluting with 5-95% acetonitrile/H₂O to afford8-((4-bromo-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(24 mg, 56%). ¹H NMR (400 MHz, CDCl₃) δ 11.08 (s, 1H), 7.30-7.27 (m,1H), 7.18-7.15 (m, 1H), 6.63 (t, 1H), 5.72 (s, 1H), 3.50-3.46 (m, 2H),3.26 (s, 3H), 2.88 (t, 2H), 2.11 (s, 3H) ppm. MS (apci, m/z)=380.0(M+H).

Example 48

8-((2-fluoro-4-iodophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

8-((4-bromo-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(Prepared according to Example 46; 20 mg, 0.053 mmol), copper(I) iodide(2.5 mg, 0.013 mmol), sodium iodide (16 mg, 0.11 mmol), and(1S,2S)-N1,N2-dimethylcyclohexane-1,2-diamine (3.7 mg, 0.026 mmol) werecombined in 1,4-dioxane (0.5 mL). The reaction was heated to 120° C. for16 hours. The reaction was then cooled to ambient temperature, quenchedwith saturated aq. NH₄Cl (2 mL), then extracted with EtOAc (2×5 mL). Thecombined organic phases were dried over MgSO₄, filtered, andconcentrated. The residue was purified by reverse phase HPLC (5-95%acetonitrile/H₂O/0.1% TFA). Fractions containing the clean desiredproduct were combined and the product was converted to the free basewith DCM/NaHCO₃ to afford8-((2-fluoro-4-iodophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(5.0 mg, 22%). ¹H NMR (400 MHz, CDCl₃) δ 11.07 (s, 1H), 7.46-7.43 (m,1H), 7.36-7.33 (m, 1H), 6.48 (t, 1H), 5.82 (s, 1H), 3.50-3.46 (m, 2H),3.27 (s, 3H), 2.88 (t, 2H), 2.11 (s, 3H) ppm. MS (apci, m/z)=428.10(M+H).

Example 49

8-((4-iodo-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionetrifluoroacetate

Step 1. Preparation of methyl(E)-4-(2-ethoxyvinyl)-2-((2-fluoro-4-iodophenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.4-iodo-2-fluoroaniline (266 mg, 1.12 mmol) and methyl(E)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(290 mg, 0.920 mmol) were dissolved in THF (9.2 mL) and cooled to −78°C. under N₂ atmosphere. LiHMDS (2.1 mL, 1.0 M/THF, 2.1 mmol) was addeddropwise, then the mixture allowed to warm from −78° C. to ambienttemperature over 1 hour. The reaction mixture was quenched withsaturated aq. NH₄Cl (10 mL), then extracted with EtOAc (2×10 mL). Thecombined organic phases were dried over MgSO₄, filtered, andconcentrated. The residue was purified by column chromatography elutingwith 0-100% EtOAc/Heptane to afford methyl(E)-4-(2-ethoxyvinyl)-2-((2-fluoro-4-iodophenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(389 mg, 77%). MS (apci, m/z)=473.1 (M+H).

Step 2. Preparation of methyl2-((4-iodo-2-fluorophenyl)amino)-1-methyl-6-oxo-4-(2-oxoethyl)-1,6-dihydropyridine-3-carboxylate.To a solution of methyl(E)-2-((4-iodo-2-fluorophenyl)amino)-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(389 mg, 0.824 mmol) in DCM (4.0 mL) was added TFA (4.0 mL). Thesolution was allowed to stir at ambient temperature for 16 hours thenconcentrated. The crude mixture was then diluted with DCM (10 mL) andwashed with saturated aq. NaHCO₃ (10 mL). The organic phase was driedover MgSO₄, filtered, and concentrated to afford methyl2-((4-iodo-2-fluorophenyl)amino)-1-methyl-6-oxo-4-(2-oxoethyl)-1,6-dihydropyridine-3-carboxylatewhich was used directly in the next step without further purification(351 mg, 96%). MS (apci, m/z)=445.0 (M+H).

Step 3. Preparation of8-((4-iodo-2-fluorophenyl)amino)-2-(4-methoxybenzyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Methyl2-((4-iodo-2-fluorophenyl)amino)-1-methyl-6-oxo-4-(2-oxoethyl)-1,6-dihydropyridine-3-carboxylate(351 mg, 0.790 mmol), (4-methoxyphenyl)methanamine (114 mg, 0.830 mmol),sodium triacetoxyborohydride (184 mg, 0.869 mmol), and acetic acid (4.8mg, 0.079 mmol) were dissolved in DCE (10 mL) and allowed to stir atambient temperature for 1 hour. At that time the reaction was warmed to60° C. and allowed to stir for 1 hour. The reaction was then allowed tocool to ambient temperature, diluted with DCM (5 mL) and washed withsaturated aq. NaHCO₃ (10 mL). The organic phase was dried over MgSO₄,filtered, and concentrated. The residue was purified by columnchromatography eluting with 0-100% EtOAc/Heptane to afford8-((4-iodo-2-fluorophenyl)amino)-2-(4-methoxybenzyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(321 mg, 76%). MS (apci, m/z)=534.1 (M+H).

Step 4. Preparation of8-((4-iodo-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionetrifluoroacetate.8-((4-iodo-2-fluorophenyl)amino)-2-(4-methoxybenzyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(321 mg, 0.602 mmol) was dissolved in TFA (6.0 mL) and heated to 60° C.for 16 hours. The reaction was then allowed to cool to ambienttemperature and concentrated. The residue was purified by reverse phaseHPLC (5-95% acetonitrile/H₂O/0.1% TFA). The fractions containing theclean desired product were lyophilized to afford8-((4-iodo-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionetrifluoroacetate (102 mg, 41%). ¹H NMR (400 MHz, CDCl₃) δ 11.27 (s, 1H),7.48-7.45 (m, 1H), 7.40-7.38 (m, 1H), 6.58 (t, 1H), 6.05 (s, 1H), 5.84(s, 1H), 3.51-3.47 (m, 2H), 3.20 (s, 3H), 2.84 (t, 2H) ppm. MS (apci,m/z)=414.0 (M+H).

Example 50

8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionetrifluoroacetate

Step 1. Preparation of methyl(E)-4-(2-ethoxyvinyl)-2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.Methyl(E)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(400 mg, 1.47 mmol) and 2-fluoro-4-(methylthio)aniline (243 mg, 1.55mmol) were dissolved in THF (9.8 mL) and cooled to −78° C. under N₂atmosphere. LiHMDS (2.9 mL, 1.0 M/THF, 2.9 mmol) was added dropwise,then the mixture allowed to warm from −78° C. to ambient temperatureover 1 hour. The reaction mixture was quenched with saturated aq. NH₄Cl(10 mL), then extracted with EtOAc (2×10 mL). The combined organicphases were dried over MgSO₄, filtered, and concentrated. The residuewas purified by column chromatography eluting with 0-100% EtOAc/heptaneto afford methyl(E)-4-(2-ethoxyvinyl)-2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(495 mg, 86%). MS (apci, m/z)=393.1 (M+H).

Step 2. Preparation of methyl2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-4-(2-oxoethyl)-1,6-dihydropyridine-3-carboxylate.To a solution of methyl(E)-4-(2-ethoxyvinyl)-2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(495 mg, 1.26 mmol) in DCM (4.0 mL) was added TFA (4.0 mL). The solutionwas allowed to stir at ambient temperature overnight then concentrated.The crude mixture was then diluted with DCM (10 mL) and washed withsaturated aq. NaHCO₃ (10 mL). The organic phase was dried over MgSO₄,filtered, and concentrated to afford methyl2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-4-(2-oxoethyl)-1,6-dihydropyridine-3-carboxylatewhich was used without further purification (391 mg, 85%). MS (apci,m/z)=365.1 (M+H).

Step 3. Preparation of8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(4-methoxybenzyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Methyl2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-4-(2-oxoethyl)-1,6-dihydropyridine-3-carboxylate(391 mg, 1.07 mmol), (4-methoxyphenyl)methanamine (155 mg, 1.13 mmol),sodium triacetoxyborohydride (250 mg, 1.18 mmol), and acetic acid (6.4mg, 0.079 mmol) were dissolved in DCE (11 mL) and allowed to stir atambient temperature for 1 hour. At that time the reaction was warmed to60° C. and allowed to stir for 1 hour. The reaction was then allowed tocool to ambient temperature, diluted with DCM (5 mL) and washed withsaturated aq. NaHCO₃ (10 mL). The organic phase was dried over MgSO₄,filtered, and concentrated. The residue was purified by columnchromatography eluting with 0-100% EtOAc/heptane to afford8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(4-methoxybenzyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(401 mg, 82%). MS (apci, m/z)=454.1 (M+H).

Step 4. Preparation of8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionetrifluoroacetate.8-((2-Fluoro-4-(methylthio)phenyl)amino)-2-(4-methoxybenzyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(401 mg, 0.884 mmol) was dissolved in TFA (8.0 mL) and heated to 60° C.for 16 hours. The reaction was then allowed to cool to ambienttemperature and concentrated. The residue was purified by reverse phaseHPLC (5-95% acetonitrile/H₂O/0.1% TFA). Fractions containing the cleandesired product were combined and lyophilized to afford8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionetrifluoroacetate (131 mg, 44%). ¹H NMR (400 MHz, CDCl₃) δ 11.10 (s, 1H),7.04-6.96 (m, 2H), 6.84 (t, 1H), 6.49 (s, 1H), 6.11 (s, 1H), 3.53-3.49(m, 2H), 3.20 (s, 3H), 2.85 (t, 2H), 2.48 (s, 3H) ppm. MS (apci,m/z)=334.1 (M+H).

Example 51

8-((4-ethyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 50, substituting 4-ethyl-2-fluoroanilinein place of 2-fluoro-4-(methylthio)aniline in step 1 to afford8-((4-ethyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(14.9 mg, 47%). ¹H NMR (400 MHz, CDCl₃) δ 11.25 (s, 1H), 6.95 (m, 1H),6.90 (m, 1H), 6.81 (t, 1H), 6.06 (s, 1H), 5.99 (s, 1H), 3.48 (m, 2H),3.17 (s, 3H), 2.82 (t, 2H), 2.62 (q, 2H), 1.22 (t, 3H) ppm; MS (apci,m/z)=316.1 (M+H).

Example 52

8-((4-cyclopropyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 50, substituting4-cyclopropyl-2-fluoroaniline in place of 2-fluoro-4-(methylthio)anilinein step 1 to afford8-((4-cyclopropyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(5.8 mg, 17%). ¹H NMR (400 MHz, CDCl₃) δ 11.24 (s, 1H), 6.83-6.75 (m,3H), 5.99 (s, 1H), 5.94 (s, 1H), 3.48 (dt, 2H), 3.16 (s, 3H), 2.82 (t,2H), 1.86 (m, 1H), 1.02-0.95 (m, 2H), 0.69-0.62 (m, 2H) ppm; MS (apci,m/z)=328.1 (M+H).

Example 53

8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 50, substituting4-(difluoromethoxy)-2-fluoroaniline in place of2-fluoro-4-(methylthio)aniline in step 1 to afford8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(30.6 mg, 47%). ¹H NMR (400 MHz, CDCl₃) δ 11.29 (s, 1H), 6.97 (m, 1H),6.91-6.84 (m, 2H), 6.49 (t, 1H), 6.03 (s, 1H), 5.79 (s, 1H), 3.49 (dt,2H), 3.19 (s, 3H), 2.84 (t, 2H) ppm; MS (apci, m/z)=354.1 (M+H).

Example 54

8-((2-fluoro-4-propylphenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Steps 1-4: Preparation of8-((4-allyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 50, substituting 4-allyl-2-fluoroanilinein place of 2-fluoro-4-(methylthio)aniline in step 1 to afford8-((4-allyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(3.8 mg, 10%). ¹H NMR (400 MHz, CDCl₃) δ 11.27 (s, 1H), 7.09 (dd, 1H),7.01 (dd, 1H), 6.80 (t, 1H), 6.35-6.28 (m, 1H), 6.25-6.15 (m, 1H), 6.01(s, 1H), 5.85 (s, 1H), 3.49 (dt, 2H), 3.19 (s, 3H), 2.83 (t, 2H), 1.88(dd, 2H) ppm; MS (apci, m/z)=328.1 (M+H).

Step 5. Preparation of8-((2-fluoro-4-propylphenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.A stirred mixture of8-((4-allyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(3.8 mg, 0.012 mmol) and Pd/C (5% Degussa type, 1 mg, 0.009 mmol) inMeOH (0.5 mL) was degassed and stirred under H₂ atmosphere for 24 hours.The reaction was diluted with DCM, filtered and the filtrateconcentrated. The residue was purified by reverse phase HPLC (5-95%acetonitrile/water/0.1% TFA over 20 minutes) and the clean fractionswere combined, washed with saturated bicarbonate, and extracted withEtOAc (3×10 mL). The combined organic layers were washed with saturatedbicarbonate (30 mL) and brine (30 mL), dried over sodium sulfate,filtered, and concentrated to afford8-((2-fluoro-4-propylphenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(2.5 mg, 65%). ¹H NMR (400 MHz, CDCl₃) δ 11.22 (s, 1H), 6.93 (dd, 1H),6.88 (m, 1H), 6.81 (t, 1H), 5.99 (s, 1H), 5.81 (s, 1H), 3.48 (dt, 2H),3.17 (s, 3H), 2.83 (t, 2H), 2.55 (t, 2H), 1.62 (m, 2H), 0.92 (t, 3H)ppm; MS (apci, m/z)=330.1 (M+H).

Example 55

8-((2-fluoro-4-(methylthio)phenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1.8-((2-fluoro-4-(methylthio)phenyl)amino)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.8-((2-Fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(41.2 mg, 0.124 mmol) and p-toluenesulfonic acid (31.9 mg, 0.185 mmol)were dissolved in 1 mL of a 1:1 THF/MeOH solution. After 5 minutes, NIS(27.8 mg, 0.124 mmol) was added. The reaction was allowed to stir atambient temperature for 30 minutes. The crude reaction mixture was thenpartitioned between EtOAc (10 mL) and saturated aq. NaHCO₃ (15 mL) andextracted with EtOAc (2×5 mL). The combined organic phases were driedover MgSO₄, filtered, and concentrated. The residue was purified bycolumn chromatography, eluting with 10-80% EtOAc/heptane to afford8-((4-bromo-2-fluorophenyl)amino)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(37 mg, 65%). MS (apci, m/z)=460.0 (M+H).

Step 2. Preparation of8-((2-fluoro-4-(methylthio)phenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.8-((2-fluoro-4-(methylthio)phenyl)amino)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(37.0 mg, 0.0806 mmol) and bis(tri-t-butylphosphine)palladium(0) (4.12mg, 0.00806 mmol) were dissolved in THF (0.8 mL) and methylzinc(II)chloride (80.6 μL, 2M/THF, 0.161 mmol) was added dropwise. The reactionwas allowed to stir at ambient temperature for 1 hour. The reactionmixture was quenched with saturated aq. NH₄Cl (2 mL), then extractedwith EtOAc (2×5 mL). The combined organic phases were dried over MgSO₄,filtered, and concentrated. The residue was purified by reverse phaseHPLC (5-95% acetonitrile/H₂O/0.1% TFA). Fractions containing the cleandesired product were combined and the product was converted to the freebase with DCM/NaHCO₃ to afford8-((2-fluoro-4-(methylthio)phenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(15.5 mg, 55%). ¹H NMR (400 MHz, CDCl₃) δ 11.10 (s, 1H), 7.04-7.01 (m,1H), 6.95-6.93 (m, 1H), 6.71 (t, 1H), 5.96 (s, 1H), 3.49-3.46 (m, 2H),3.25 (s, 3H), 2.87 (t, 2H), 2.46 (s, 3H), 2.11 (s, 3H) ppm. MS (apci,m/z)=348.1 (M+H).

Example 56

8-((4-ethyl-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 55, substituting8-((4-ethyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(prepared according to Example 51) in place of8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionein step 1 to afford8-((4-ethyl-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(6.7 mg, 62%). ¹H NMR (400 MHz, CDCl₃) δ 11.06 (s, 1H), 6.94 (dd, 1H),6.86 (dd, 1H), 6.72 (t, 1H), 5.87 (s, 1H), 3.47 (dt, 2H), 3.23 (s, 3H),2.87 (t, 2H), 2.61 (q, 2H), 2.11 (s, 3H), 1.21 (t, 3H) ppm; MS (apci,m/z)=330.2 (M+H).

Example 57

8-((4-cyclopropyl-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 55, substituting8-((4-cyclopropyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(prepared according to Example 52) in place of8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionein step 1 to afford8-((4-cyclopropyl-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(2.3 mg, 47%). ¹H NMR (400 MHz, CDCl₃) δ 11.05 (s, 1H), 6.80-6.75 (m,2H), 6.69 (t, 1H), 5.80 (s, 1H), 3.47 (dt, 2H), 3.22 (s, 3H), 2.87 (t,2H), 2.10 (s, 3H), 1.85 (m, 1H), 1.00-0.94 (m, 2H), 0.67-0.61 (m, 2H)ppm; MS (apci, m/z)=342.2 (M+H).

Example 58

8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 55, substituting8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(prepared according to Example 53) in place of8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionein step 1 to afford8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(9.5 mg, 41%). ¹H NMR (400 MHz, CDCl₃) δ 11.09 (s, 1H), 6.96 (dd, 1H),6.87-6.82 (m, 1H), 6.76 (t, 1H), 6.47 (t, 1H), 6.01 (s, 1H), 3.48 (dt,2H), 3.25 (s, 3H), 2.88 (t, 2H), 2.11 (s, 3H) ppm; MS (apci, m/z)=368.1(M+H).

Example 59

8-((4-bromo-2-chlorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of methyl(E)-2-((4-bromo-2-chlorophenyl)amino)-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.Methyl(E)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(140 mg, 0.515 mmol) and 4-bromo-2-chloroaniline (108 mg, 0.520 mmol)were dissolved in THF (5.2 mL) and cooled to −78° C. under N₂atmosphere. LiHMDS (1.0 mL, 1.0 M/THF, 1.0 mmol) was added dropwise,then the mixture allowed to warm from −78° C. to ambient temperatureover 1 hour. The reaction mixture was quenched with saturated aq. NH₄Cl(5 mL), then extracted with EtOAc (2×5 mL). The combined organic phaseswere dried over MgSO₄, filtered, and concentrated. The residue waspurified by column chromatography eluting with 0-100% EtOAc/Heptane toafford methyl(E)-2-((4-bromo-2-chlorophenyl)amino)-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(145 mg, 64%). MS (apci, m/z)=441.0, 443.0 (M+H).

Step 2. Preparation of methyl2-((4-bromo-2-chlorophenyl)amino)-1-methyl-6-oxo-4-(2-oxoethyl)-1,6-dihydropyridine-3-carboxylate.To a solution of methyl(E)-2-((4-bromo-2-chlorophenyl)amino)-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(145 mg, 0.328 mmol) in DCM (2.0 mL) was added TFA (2.0 mL). Thesolution was allowed to stir at ambient temperature overnight, thenconcentrated. The crude mixture was then diluted with DCM (5 mL) andwashed with saturated aq. NaHCO₃ (5 mL). The organic phase was driedover MgSO₄, filtered, and concentrated to afford methyl2-((4-bromo-2-chlorophenyl)amino)-1-methyl-6-oxo-4-(2-oxoethyl)-1,6-dihydropyridine-3-carboxylatewhich was used without further purification (136 mg, 100%). MS (apci,m/z)=415.2 (M+H).

Step 3. Preparation of8-((4-bromo-2-chlorophenyl)amino)-2-(4-methoxybenzyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Methyl2-((4-bromo-2-chlorophenyl)amino)-1-methyl-6-oxo-4-(2-oxoethyl)-1,6-dihydropyridine-3-carboxylate(136 mg, 0.329 mmol), (4-methoxyphenyl)methanamine (47.4 mg, 0.345mmol), sodium triacetoxyborohydride (76.6 mg, 0.362 mmol), and aceticacid (2.0 mg, 0.033 mmol) were dissolved in DCE (3.3 mL) and allowed tostir at ambient temperature for 1 hour. At that time the reaction waswarmed to 60° C. and allowed to stir for 1 hour. The reaction was thenallowed to cool to ambient temperature, diluted with DCM (5 mL) andwashed with saturated aq. NaHCO₃ (10 mL). The organic phase was driedover MgSO₄, filtered, and concentrated. The residue was purified bycolumn chromatography eluting with 0-100% EtOAc/Heptane to afford8-((4-bromo-2-chlorophenyl)amino)-2-(4-methoxybenzyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(104 mg, 63%). MS (apci, m/z)=502.1, 504.1 (M+H).

Step 4. Preparation of8-((4-bromo-2-chlorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.8-((4-bromo-2-chlorophenyl)amino)-2-(4-methoxybenzyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(79.1 mg, 0.207 mmol) was dissolved in TFA (3.0 mL) and heated to 60° C.for 16 hours. The reaction was then allowed to cool to ambienttemperature and concentrated. The residue was purified by reverse phaseHPLC (5-95% acetonitrile/H₂O/0.1% TFA). Fractions containing the cleandesired product were combined and the product was converted to the freebase with DCM/NaHCO₃ to afford8-((4-bromo-2-chlorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(28 mg, 35%). 1H NMR (400 MHz, CDCl₃) δ 11.27 (s, 1H), 7.61 (d, 1H),7.31-7.28 (m, 1H), 6.60-6.57 (m, 1H), 6.07 (t, 1H), 5.63 (s, 1H),3.52-3.48 (m, 2H), 3.17 (s, 3H), 2.85 (t, 2H) ppm. MS (apci, m/z)=382.0,384.0 (M+H).

Example 60

8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionetrifluoroacetate

Step 1. Preparation of methyl2-chloro-1-methyl-6-oxo-4-(2-oxoethyl)-1,6-dihydropyridine-3-carboxylate.To a solution of methyl(E)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(331 mg, 1.22 mmol) in DCM (5.0 mL) was added TFA (5.0 mL). The solutionwas allowed to stir at ambient temperature for 16 hours, thenconcentrated. The crude mixture was then diluted with DCM (10 mL) andwashed with saturated aq. NaHCO₃ (10 mL). The organic phase was driedover MgSO₄, filtered, and concentrated to afford methyl2-chloro-1-methyl-6-oxo-4-(2-oxoethyl)-1,6-dihydropyridine-3-carboxylatewhich was used without further purification (236 mg, 80%). MS (apci,m/z)=244.1 (M+H).

Step 2. Preparation of8-chloro-2-(2-hydroxyethyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Methyl2-chloro-1-methyl-6-oxo-4-(2-oxoethyl)-1,6-dihydropyridine-3-carboxylate(122 mg, 0.501 mmol), 2-((tert-butyldimethylsilyl)oxy)ethan-1-amine (105mg, 0.601 mmol), sodium triacetoxyborohydride (127 mg, 0.601 mmol), andacetic acid (6.0 mg, 0.10 mmol) were dissolved in DCE (5 mL) and allowedto stir at ambient temperature for 2 hours. The reaction was warmed to60° C. and stirred for 16 hours. The reaction was then allowed to coolto ambient temperature, diluted with DCM (10 mL) and washed withsaturated aq. NaHCO₃ (15 mL). The organic phase was dried over MgSO₄,filtered, and concentrated. The residue was purified by reverse phaseHPLC (5-95% acetonitrile/H₂O/0.1% TFA). Fractions containing the cleandesired product were combined and the product was converted to the freebase with DCM/NaHCO₃ to afford8-chloro-2-(2-hydroxyethyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(20 mg, 16%). MS (apci, m/z)=257.0 (M+H).

Step 3. Preparation of8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionetrifluoroacetate. 4-Bromo-2-fluoroaniline (17.7 mg, 0.093 mmol) and8-chloro-2-(2-hydroxyethyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(20 mg, 0.077 mmol) were dissolved in THF (0.7 mL) and cooled to −78° C.under N₂ atmosphere. LiHMDS (233 μL, 1.0 M/THF, 0.233 mmol) was addeddropwise, then the mixture allowed to warm from −78° C. to ambienttemperature over 1 hour. The reaction mixture was quenched withsaturated NH₄Cl (5 mL), then extracted with EtOAc (3×5 mL). The combinedorganic phases were dried over MgSO₄, filtered, and concentrated. Theresidue was purified by reverse phase HPLC (5-95% acetonitrile/H₂O/0.1%TFA). Clean fractions were lyophilized to afford8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionetrifluoroacetate (10.1 mg, 32%). ¹H NMR (400 MHz, CDCl₃) δ 11.36 (s,1H), 7.32-7.29 (m, 1H), 7.22-7.19 (m, 1H), 6.70 (t, 1H), 6.06 (s, 1H),3.85 (t, 2H), 3.67 (t, 2H), 3.61 (t, 2H), 3.22 (s, 3H), 2.86 (t, 2H)ppm. MS (apci, m/z)=410.1, 412.1 (M+H).

Example 61

8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 44 substituting2-fluoro-4-(methylthio)aniline in place of 4-ethyl-2-fluoroaniline instep 1 to afford8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(2.1 mg, 21%). ¹H NMR (400 MHz, CDCl₃) δ 11.01 (s, 1H), 7.02 (dd, 1H),6.95 (m, 1H), 6.77 (m, 1), 4.48 (t, 1H), 4.03 (t, 2H), 3.76-3.67 (m,4H), 3.21 (s, 3H), 3.02 (t, 2H), 2.47 (s, 3H), 2.08 (s, 3H) ppm. MS(apci, m/z)=408.1 (M+H).

Example 62

8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Steps 1-5. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.Prepared according to Example 41, Steps 1-5 substituting4-(difluoromethoxy)-2-fluoroaniline in place of 2-fluoroaniline in step1 to afford2-(2-(tert-butoxy)ethoxy)-8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(156 mg, 94%). MS (apci, m/z)=596.1 (M+H).

Step 6. Preparation of2-(2-(tert-butoxy)ethoxy)-8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of2-(2-(tert-butoxy)ethoxy)-8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(156 mg, 0.262 mmol) in THF (2.5 mL) at 0° C. was added Pd(Pt-Bu₃)₂(13.4mg, 0.026 mmol) followed by methylzinc(II) chloride (132 μL, 0.265mmol). The mixture was removed from the ice bath and stirred for 10minutes. The reaction was partitioned between saturated NH₄Cl (40 mL)and EtOAc (20 mL) and the aqueous layer was extracted with EtOAc (2×10mL). The combined organic phases were washed with brine (30 mL), driedover sodium sulfate, filtered, and concentrated. The residue waspurified by column chromatography eluting with 0-50% EtOAc/heptane toafford2-(2-(tert-butoxy)ethoxy)-8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(89.9 mg, 71%). MS (apci, m/z)=484.2 (M+H).

Step 7. Preparation of8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.2-(2-(tert-butoxy)ethoxy)-8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(89.9 mg, 0.186 mmol) was dissolved in ACN (750 μL) and phosphoric acid(750 μL) was added. The reaction was heated to 60° C. for 30 minutes.The reaction mixture was partitioned between saturated sodiumbicarbonate (50 mL) and EtOAc (20 mL). The aqueous layer was washed withEtOAc (2×15 mL). The combined organic layers were washed with saturatedsodium bicarbonate (25 mL), brine (25 mL), dried over sodium sulfate,filtered, and concentrated. The residue was purified by columnchromatography eluting with 0-100% EtOAc/heptane to afford8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(65.9 mg, 83%). ¹H NMR (400 MHz, CDCl₃) δ 11.00 (s, 1H), 6.97 (dd, 1H),6.90-6.79 (m, 2H), 6.48 (t, 1H), 4.44 (t, 1H), 4.03 (m, 2H), 3.76-3.68(m, 4H), 3.22 (s, 3H), 3.03 (t, 2H), 2.09 (s, 3H) ppm. MS (apci,m/z)=428.1 (M+H).

Example 63

2-(2,2-difluoroethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of methyl(Z)-4-(2-ethoxyvinyl)-2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.Prepared according to Example 44, Step 1 substituting2-fluoro-4-(methylthio)aniline in place of 4-ethyl-2-fluoroaniline toafford methyl(Z)-4-(2-ethoxyvinyl)-2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(0.373 g, 69%). MS (apci, m/z)=393.1 (M+H).

Step 2. Preparation of methyl(E)-4-(2-((2,2-difluoroethoxy)imino)ethyl)-2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.Prepared according to Example 6, Step 5 substituting methyl(Z)-4-(2-ethoxyvinyl)-2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylatein place of methyl(Z)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylateand O-(2,2-difluoroethyl)hydroxylamine hydrochloride in place ofO-(2-(tert-butoxy)ethyl)hydroxylamine hydrochloride to afford methyl(E)-4-(2-((2,2-difluoroethoxy)imino)ethyl)-2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(assumed 100%). MS (apci, m/z)=444.1 (M+H).

Step 3. Preparation of2-(2,2-difluoroethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of methyl(E)-4-(2-((2,2-difluoroethoxy)imino)ethyl)-2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(36 mg, 0.081 mmol) in MeOH (0.8 mL) was added sodium cyanoborohydride(26 mg, 0.406 mmol) and acetic acid (23 μL, 0.406 mmol). The mixture wasstirred at 45° C. for 2 hours, then at ambient temperature for 72 hours.The mixture was diluted with sat. aqueous NaHCO₃ (25 mL) and EtOAc (25mL). The organic phase was separated, dried over Na₂SO₄, filtered, andconcentrated. The residue was purified by column chromatography, elutingwith 5-100% EtOAc/heptane, to afford2-(2,2-difluoroethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(11.8 mg, 35.2%). ¹H NMR (400 MHz, CDCl₃) δ 11.26 (s, 1H), 7.08-6.94 (m,2H), 6.91-6.76 (m, 1H), 6.51-5.64 (m, 2H), 4.22 (td, 2H), 3.73 (t, 2H),3.16 (s, 3H), 3.04-2.97 (m, 2H), 2.48 (s, 3H) ppm. MS (apci, m/z)=414.1(M+H).

Example 64

8-((4-bromo-2-fluorophenyl)amino)-7-methyl-4-phenyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of 2,6-dichloro-4-iodonicotinic acid. A solution of2,6-dichloro-4-iodopyridine (4.0 g, 14.6 mmol) in THF (30 mL) was cooledto −78° C. LDA (10.95 mL, 2M, 21.9 mmol) was added dropwise to thesolution at −78° C. and stirred 2 hours at −78° C. Excess CO₂ gas wasbubbled into the reaction solution for 20 minutes at −78° C. Thereaction mixture was then poured into dry ice (10 g) and quenched withwater (30 mL). The reaction was adjusted to pH 3-4 with 2N HCl andextracted with EtOAc (3×100 mL). The organic layers were combined anddried over sodium sulfate, filtered, and concentrated to afford2,6-dichloro-4-iodonicotinic acid (3.0 g, 64.6%). MS (apci, m/z)=317.9,319.8 (M+H).

Step 2. Preparation of2-chloro-4-iodo-6-oxo-1,6-dihydropyridine-3-carboxylic acid. A solutionof NaOH (100 mL, 4M, 236 mmol) was heated to 110° C. and then2,6-dichloro-4-iodonicotinic acid (3.0 g, 9.44 mmol) was added in oneportion and stirred for 8 hours. The reaction was adjusted pH to 1 withHCl (6 M) at 0° C. and stirred for 30 minutes. The resulting solids werecollected by filtration and dried in vacuo to afford2-chloro-4-iodo-6-oxo-1,6-dihydropyridine-3-carboxylic acid (3.0 g,assumed 100%) which was used without further purification. MS (apci,m/z) 20=299.9, 301.9 (M+H).

Step 3. Preparation of methyl2-chloro-4-iodo-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate. To amixture of 2-chloro-4-iodo-6-oxo-1,6-dihydropyridine-3-carboxylic acid(3.0 g, 10.02 mmol) in DMF (30 mL) was added Mel (4.27 g, 30.06 mmol)and K₂CO₃ (4.15 g, 30.06 mmol) in one portion at 25° C. After stirringfor 3 hours, the reaction was poured into sat. NH₄Cl (10 mL) and theaqueous phase was extracted with ethyl acetate (3×50 mL). The combinedorganic phases were washed with brine (100 mL), dried over Na₂SO₄,filtered and concentrated. The residue was purified by columnchromatography eluting with 2-100% EtOAc/petroleum ether to affordmethyl 2-chloro-4-iodo-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(850 mg, 25.9%). ¹H NMR (400 MHz, CDCl₃) δ 7.16 (s, 1H), 3.93 (s, 3H),3.65 (s, 3H) ppm. MS (apci, m/z)=327.9 (M+H).

Step 4. Preparation of methyl2-chloro-1-methyl-6-oxo-4-(1-phenylvinyl)-1,6-dihydropyridine-3-carboxylate.To a solution of methyl2-chloro-4-iodo-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate (0.2 g,0.611 mmol) in 1,4-dioxane (10 mL) and water (1 mL) was added4,4,5,5-tetramethyl-2-(1-phenylvinyl)-1,3,2-dioxaborolane (0.148 g,0.641 mmol), Pd(dppf)Cl₂ (50 mg, 0.061 mmol), Na₂CO₃ (0.194 g, 1.83mmol) and H₂O (1 mL). Then the mixture was degassed 3 times and stirredat 80° C. for 3 hours.4,4,5,5-Tetramethyl-2-(1-phenylvinyl)-1,3,2-dioxaborolane (37 mg, 0.153mmol) and Pd(dppf)Cl₂ (50 mg, 0.061 mmol) were added. The mixture wasdegassed 3 times and stirred at 80° C. for another 1.5 hours. The cooledmixture was filtered and the filtrate concentrated. The residue waspurified by column chromatography eluting with 0-20% EtOAc/petroleumether to afford methyl2-chloro-1-methyl-6-oxo-4-(1-phenylvinyl)-1,6-dihydropyridine-3-carboxylate(95 mg, 51%). ¹H NMR (400 MHz, CDCl₃) δ 7.33-7.32 (m, 3H), 7.32-7.25 (m,2H), 6.58 (s, 1H), 5.62 (s, 1H), 5.42 (s, 1H), 3.75 (s, 3H), 3.36 (s,3H) ppm. MS (apci, m/z)=303.7 (M+H).

Step 5. Preparation of methyl2-((4-bromo-2-fluorophenyl)amino)-1-methyl-6-oxo-4-(1-phenylvinyl)-1,6-dihydropyridine-3-carboxylate.To a solution of 4-bromo-2-fluoroaniline (56 mg, 0.29 mmol) in THF (5mL) was added LiHMDS (0.74 mL, 1.0 M, 0.74 mmol) at −78° C. and stirredfor 0.5 hours at −78° C. Then a solution of methyl2-chloro-1-methyl-6-oxo-4-(1-phenylvinyl)-1,6-dihydropyridine-3-carboxylate(90 mg, 0.296 mmol) in THF (0.5 mL) was added and the mixture wasstirred at −78° C. for 30 minutes. The mixture was quenched withsaturated NH₄Cl solution (5 mL) and extracted with EtOAc (3×50 mL). Thecombined organic phases were washed with brine (5 mL), dried overNa₂SO₄, filtered and concentrated. The residue was purified by columnchromatography eluting with 0-20% EtOAc/petroleum ether to afford methyl2-((4-bromo-2-fluorophenyl)amino)-1-methyl-6-oxo-4-(1-phenylvinyl)-1,6-dihydropyridine-3-carboxylate(120 mg, 89%). MS (apci, m/z)=459.2 (M+H).

Step 6. Preparation of8-((4-bromo-2-fluorophenyl)amino)-2-(2,4-dimethoxybenzyl)-7-methyl-4-phenyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of methyl2-((4-bromo-2-fluorophenyl)amino)-1-methyl-6-oxo-4-(1-phenylvinyl)-1,6-dihydropyridine-3-carboxylate(120 mg, 0.29 mmol) in toluene (10 mL) was added DMB-NH₂ (43.8 mg, 0.26mmol) and AlMe₃(0.4 mL, 2.0 M, 0.79 mmol) at 25° C. under N₂ and themixture was stirred at 80° C. for 48 hours. The mixture was quenchedwith saturated NH₄Cl solution (5 mL) and filtered. The filtrate wasextracted with EtOAc (3×10 mL) and the organic layer was washed withbrine (10 mL), dried over Na₂SO₄, filtered and concentrated. The residuewas purified by column chromatography eluting with 0-50% EtOAc/petroleumether to afford8-((4-bromo-2-fluorophenyl)amino)-2-(2,4-dimethoxybenzyl)-7-methyl-4-phenyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(80.0 mg, 52%). MS (apci, m/z)=594.2 (M+H).

Step 7.8-((4-bromo-2-fluorophenyl)amino)-7-methyl-4-phenyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.A mixture of8-((4-bromo-2-fluorophenyl)amino)-2-(2,4-dimethoxybenzyl)-7-methyl-4-phenyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(80 mg, 0.14 mmol) in TFA (5 mL) was stirred for 1 hour at 80° C. Thecooled mixture was concentrated and the residue was dissolved in MeOH (5mL). NaHCO₃ solid (˜1 g) was added and the mixture stirred for 15minutes then treated with DCM (20 mL) and filtered. The filtrate wasconcentrated and purified by prep HPLC (Prep HPLC condition: column:Boston Prime C18 150*25 mm*5 um; mobile phase: water (0.225% ammoniahydroxide v/v)-ACN; B %: 40%-70%, FlowRate (ml/min): 25). Fractions werelyophilized to afford8-((4-bromo-2-fluorophenyl)amino)-7-methyl-4-phenyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(5.49 mg, 9%). ¹H NMR (400 MHz, MeOD) δ 7.49 (d, 1H), 7.40-7.32 (m, 6H),6.93 (t, 1H), 5.76 (s, 1H), 4.19 (t, 1H), 3.66 (d, 2H), 3.24 (s, 3H)ppm. MS (apci, m/z)=442.2, 444.1 (M+H).

Example 65

8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-4-phenyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 64, substituting2-fluoro-4-(methylthio)aniline in place of 4-bromo-2-fluoroaniline instep 5 to afford8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-4-phenyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(21 mg, 29%). ¹H NMR (400 MHz, d₆-DMSO) δ 11.71 (s, 1H), 8.15 (s, 1H),7.40-7.25 (m, 6H), 7.08 (d, 1H), 6.97 (t, 1H), 5.59 (s, 1H), 4.13 (t,1H), 3.54 (bs, 2H), 3.30 (s, 3H), 3.05 (s, 3H) ppm. MS (apci, m/z)=410.1(M+H).

Example 66

8-((4-bromo-2-fluorophenyl)amino)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 64, substituting4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane in place of4,4,5,5-tetramethyl-2-(1-phenylvinyl)-1,3,2-dioxaborolane in Step 4 toafford8-((4-bromo-2-fluorophenyl)amino)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(3 mg, 13%). ¹H NMR (400 MHz, MeOD) δ 7.48 (d, 1H), 7.35 (d, 1H), 6.88(t, 1H), 6.15 (s, 1H), 3.51 (dd, 1H), 3.21 (s, 3H), 3.20-3.15 (m, 1H),2.99-2.86 (m, 1H), 1.33 (d, 3H) ppm. MS (apci, m/z)=380.0, 382.2 (M+H).

Example 67

8-((2-fluoro-4-(methylthio)phenyl)amino)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionePrepared according to Example 64, substituting4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane in place of4,4,5,5-tetramethyl-2-(1-phenylvinyl)-1,3,2-dioxaborolane in Step 4 and2-fluoro-4-(methylthio)aniline in place of 4-bromo-2-fluoroaniline instep 5 to afford8-((2-fluoro-4-(methylthio)phenyl)amino)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(3.8 mg, 22%). ¹H NMR (400 MHz, MeOD) δ 7.16 (d, 1H), 7.09 (d, 1H), 6.92(t, 1H), 6.09 (s, 1H), 3.51 (dd, 1H), 3.19 (s, 3H), 3.17-3.15 (m, 1H),2.99-2.94 (m, 1H), 2.51 (s, 3H), 1.33 (d, 3H) ppm. MS (apci, m/z)=348.1(M+H).

Example 68

8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Step 1. Preparation of 1-ethoxyprop-1-yne. Anhydrous NH₃ (300 mL) wascondensed at −70° C. in a reaction flask, and crushed Fe(NO₃)₃-9H₂O (252mg, 0.625 mmol) was added with slow stirring. Solid NaNH₂ (34.14 g,0.875 mol) was added at −60 to −40° C. over 10 minutes, then2-chloro-1,1-diethoxyethane (38.15 g, 0.25 mol) was added dropwise atthis temperature over a period of 30 minutes. The reaction was refluxedat −30° C. for 1 hour, prior to dropwise addition of Mel (177.42 g, 1.25mol) over 30 minutes. The reaction mixture was stirred vigorously for 90minutes at this temperature, then cautiously quenched by dropwiseaddition of a cooled sat. NH₄Cl solution (40 mL), followed by Et₂O (100mL) and additional sat. NH₄Cl (50 mL). The cooling bath was removed, andthe reaction mixture was allowed to warm to 15° C. Water (50 mL) wasadded, and the organic layer was separated and washed with water (50 mL)and brine (50 mL). The combined aqueous solutions were extracted withEt₂O (50 mL), and the combined organic layers were dried over Na₂SO₄ andfiltered to afford ˜1.3 M ethereal solution of 1-ethoxyprop-1-yne (100mL, 52%). ¹H NMR (400 MHz, CDCl₃) δ 3.99 (q, 2H), 1.73 (s, 3H), 1.32 (t,3H) ppm.

Step 2. Preparation of(Z)-2-(1-ethoxyprop-1-en-2-yl)benzo[d][1,3,2]dioxaborole. To a solutionof benzo[d][1,3,2]dioxaborole (1.43 g, 11.89 mmol) in toluene (50 mL)was added 1-ethoxyprop-1-yne (7.7 mL, 1.3M/Et₂O, 11.89 mmol) andNiCl₂(dppe) (314 mg, 0.59 mmol) under N₂. The reaction mixture wasdegassed three times and stirred at 50° C. for 3 hours. The mixture wasfiltered to afford a toluene solution of(Z)-2-(1-ethoxyprop-1-en-2-yl)benzo[d][1,3,2]dioxaborole (1.5 g, 62%)which was used without further purification.

Step 3. Preparation of methyl(E)-2-chloro-4-(1-ethoxyprop-1-en-2-yl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.To a solution of(Z)-2-(1-ethoxyprop-1-en-2-yl)benzo[d][1,3,2]dioxaborole (1.5 g, 7.33mmol) in toluene (50 mL) and THF (30 mL) was added methyl2-chloro-4-iodo-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate (1.2 g,3.66 mmol), Pd(dppf)Cl₂ (299 mg, 0.37 mmol), K₃PO₄ (2.33 g, 10.99 mmol)and H₂O (3 mL). The mixture was degassed 3 times and stirred at 75° C.for 3 hours. The cooled mixture was filtered. The filtrate was dilutedwith EtOAc (50 mL) then washed with water (3×10 mL) and brine (10 mL),dried over Na₂SO₄, filtered and concentrated. The residue was purifiedby column chromatography eluting with 0-40% EtOAc/petroleum ether toafford methyl(E)-2-chloro-4-(1-ethoxyprop-1-en-2-yl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(650 mg, 62%). MS (apci, m/z)=286.0 (M+H).

Step 4. Preparation of methyl(E/Z)-4-(1-((2-(tert-butoxy)ethoxy)imino)propan-2-yl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate.To a solution of methyl(E)-2-chloro-4-(1-ethoxyprop-1-en-2-yl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(600 mg, 2.1 mmol) and O-(2-(tert-butoxy)ethyl)hydroxylaminehydrochloride (356 mg, 2.1 mmol) in dioxane (10 mL) were added TEA (212mg, 2.1 mmol) and HCl (1 mL, 4M/dioxane, 4.2 mmol) The mixture washeated to 60° C. and stirred for 2 hours. 1,4-Dioxane (20 mL) was addedand the mixture was filtered and concentrated to afford methyl(E/Z)-4-(1-((2-(tert-butoxy)ethoxy)imino)propan-2-yl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(780 mg, 99.6%) which was used without further purification. MS (apci,m/z)=317.1 (M+H−t-Bu).

Step 5. Preparation of2-(2-(tert-butoxy)ethoxy)-8-chloro-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of methyl(E/Z)-4-(1-((2-(tert-butoxy)ethoxy)imino)propan-2-yl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(780 mg, 2.09 mmol) in IPA (10 mL) were added NaCNBH₃ (657 mg, 10.46mmol) and AcOH (628 mg, 10.46 mmol). The mixture was stirred at 20° C.for 48 hours. EtOAc (100 mL) was added and the mixture was washed withsaturated NaHCO₃ (10 mL) and brine (10 mL), dried over MgSO₄, filteredand concentrated. The residue was purified by column chromatographyeluting with 0-40% EtOAc/petroleum ether to afford2-(2-(tert-butoxy)ethoxy)-8-chloro-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(200 mg, 28%). ¹H NMR (400 MHz, CDCl₃) δ 6.31 (s, 1H), 4.10-4.06 (m,2H), 3.84 (dd, 1H), 3.70 (s, 3H), 3.59-3.53 (m, 3H), 3.06-3.05 (m, 1H),1.31 (d, 3H), 1.15 (s, 9H) ppm. MS (apci, m/z)=343.1 (M+H).

Step 6. Preparation of8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.To a solution of 4-bromo-2-fluoroaniline (55 mg, 0.29 mmol) in THF (8mL) was added LiHMDS (0.73 mL, 1M, 0.73 mmol) dropwise at −78° C. underN₂. The mixture was stirred at −78° C. for 30 minutes then a solution of2-(2-(tert-butoxy)ethoxy)-8-chloro-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(100 mg, 0.29 mmol) in THF (2 mL) was added dropwise and stirringcontinued at −78° C. for 30 minutes. The mixture was treated with sat.NH₄Cl (5 mL) and extracted with EtOAc (2×30 mL). The combined organiclayers were washed with brine (5 mL), dried over MgSO₄, filtered andconcentrated to afford8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(144.8 mg, 100%) which was used without further purification. MS (apci,m/z)=496.2 (M+H).

Step 7. Preparation of8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.A mixture of8-((4-bromo-2-fluorophenyl)amino)-2-(2-(tert-butoxy)ethoxy)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(144.8 mg, 0.292 mmol) in TFA (5 mL) was stirred for 2 hours at 25° C.The mixture was concentrated, dissolved in MeOH (10 mL) and treated withNaHCO₃ solid (73 mg, 0.876 mmol). After stirring for 30 minutes, DCM (50mL) was added. The mixture was filtered, concentrated and purified byprep HPLC (Prep HPLC condition: column: Waters xbridge 150*25 mm 10 um;mobile phase: water (0.05% ammonia hydroxide v/v)-ACN; B %:24%-64%,FlowRate (ml/min): 25). Clean fractions were lyophilized to afford8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(34.8 mg, 27%). ¹H NMR (400 MHz, MeOD) δ 7.38 (d, 1H), 7.26 (d, 1H),6.82 (t, 1H), 6.02 (s, 1H), 3.98 (t, 2H), 3.82 (dd, 1H), 3.66 (t, 2H),3.64-3.50 (m, 1H), 3.12 (s, 3H), 3.09-3.08 (m, 1H), 1.29 (d, 3H) ppm. MS(apci, m/z)=440.1 (M+H).

Example 69

8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione

Prepared according to Example 68, substituting2-fluoro-4-(methylthio)aniline in place of 4-bromo-2-fluoroaniline instep 6 to afford8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione(27.5 mg, 23%). ¹H NMR (400 MHz, MeOD) δ 7.16 (d, 1H), 7.10 (d, 1H),6.99 (t, 1H), 6.07 (s, 1H), 4.05 (t, 2H), 3.91 (dd, 1H), 3.76 (t, 2H),3.60 (dd, 1H), 3.21 (s, 3H), 3.19-3.15 (m, 1H), 2.50 (s, 3H), 1.40 (d,3H) ppm. MS (apci, m/z)=408.2 (M+H).

Example 70 Preparation of anhydrous crystalline8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 1

Phosphoric acid (37.4 g, 25.8 mL, 14.8 molar, 50 Eq, 382 mmol) was addedto a stirred solution of2-(2-(tert-butoxy)ethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,which may be prepared according to any of the methods disclosed herein(3.43 g, 1 Eq, 7.63 mmol) in acetonitrile (ACN) under Air. The mixturewas warmed to 60° C. and stirred for 15 minutes after which LCMSindicated complete reaction. The reaction was cooled to 0° C. andtreated with K₃PO₄ (382 mL, 1M, 382 mmol). The mixture was extractedwith ethyl acetate (3×100 mL). The combined organic layers were washedwith brine (50 mL), dried over sodium sulfate, filtered, andconcentrated in vacuo to afford a bright yellow foam. The crude residuewas purified over 80 g silica cartridge, eluting with a gradient of 0%to 15% methanol in dichloromethane (DCM) at first, then increased to 20%methanol in DCM to afford the product as a bright yellow foam (2.17 g).This was dissolved in DCM (50 mL), treated with Norit CA1 activatedcharcoal (600 mg) and stirred for 15 min. The mixture was filteredthrough GF paper and the residue concentrated to afford a bright yellowfoam. This procedure was repeated using Darco G-60 activated charcoal toafford the product as a pale pink foam. This material was treated with2-propanol (20 mL) and DCM (5 mL) added to fully solubilize. The mixturewas concentrated, during which solids began to form but material becamean oil once fully concentrated. The residue was treated with 2-propanol(10 mL) and warmed to 40° C. upon which a thick precipitate appeared.The mixture cooled to room temperature and diluted with cold 2-propanol(5 mL) to facilitate stirring. The mixture was then filtered and 15 mLof cold 2-propanol used for rinsing and washing to afford crystallineanhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 1 (1.61 g, 4.09 mmol, 53.6%) as an off-white solid after drying invacuo. FIG. 1 depicts a powder X-ray diffraction pattern of crystallineanhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 1 prepared according to this method, wherein the PXRD analysis wasconducted using the Instrument Methods described in Example 77.

Example 71 Preparation of seed crystals of crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2

Step 1: Methyl4-bromo-2-((2-fluoro-4-(methylthio)phenyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(prepared according to any of the methods described herein) (1.0 equiv.7.5 kg), XPhos (0.075 equiv) and MeThF (10 volumes) were added to areactor and the reaction was degassed. Pd (II) acetate (0.015 equiv.)was charged to the reactor and reaction was degassed.(Z)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (0.8equiv) and sparged 45 wt % KOH solution (4 equiv.) were charged to thereactor. The reaction mixture was heated at 50° C. for 1 hour beforeadditional (Z)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(0.3 equiv) was added, and heating was continued for another hour. Thereaction mixture was washed with water (2 volumes) followed by 1M aceticacid (2 equiv.), then stirred with SiliaMetS® thiol (SiliCycle Inc.,Quebec City, Quebec, Canada) (0.3 g/g) at 50° C. for 18 hours beforefiltering. The filter cake was washed with MeTHF (5 volumes). Theorganic layer was concentrated to 3 volumes and a small portion wasremoved and cooled to generate seed crystals. The seed crystals werecharged back to the reactor and allowed to stir for 1 hour at 50° C.before slowly cooling to 30° C. MeTHF/heptane 1:4 (5 volumes) was thenslowly charged to the reactor and the mixture was stirred for 1 hour.the mixture was then slowly cooled to 10° C. and allowed to stir for 18hours before filtering. The filter cake was washed with MeTHF/heptane1:1 (5 volumes). The solids were then dried in the vacuum oven to affordmethyl4-[(Z)-2-ethoxyethenyl]-2-[2-fluoro-4-(methylsulfanyl)anilino]-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate

Step 2: Methyl4-[(Z)-2-ethoxyethenyl]-2-[2-fluoro-4-(methylsulfanyl)anilino]-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate(5.0 g, 12.5 mmol) was combined with 2-methyltetrahydrofuran (20volumes) and O-(2-(tert-butoxy)ethyl)hydroxylamine hydrochloride (1.3equiv., 16.3 mmol) and heated to approximately 75° C. The resultingmixture cooled to 20° C. Borane-pyridine complex (2.0 eq, 25.0 mmol) wasadded as well as HCl in cyclopentyl methyl ether (CPME) (3M, 1.3 eq,16.3 mmol). The resulting mixture was stirred at 20° C. for 45 min thenheated to 60° C. After stirring overnight, the mixture was cooled to 20°C., and quenched with HCl (1M, 30 mL). During this time, some bubblingand exotherm was observed. After the off gassing appeared to becomplete, (˜5 minutes), the organic layer was separated, washed withwater (8 vol), and concentrated to approximately 6 volumes via reducedpressure distillation. At this time, phosphoric acid (14.6M, 15 eq, 187mmol) was added and the mixture stirred at 65° C. After stirringovernight, the mixture was cooled to 20° C. and diluted with2-methyltetrahydrofuran (7 vol). A solution of potassium hydroxide(11.5M, 20 eq) was added slowly. After stirring for approximately 10minutes, the layers were allowed to separate. The organic phase waswashed with water (10 volumes) before concentrating to approximately 5volumes with reduced pressure distillation to afford a solution of8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dionein 2-methyltetrahydrofuran. Cyclopentyl methyl ether (2 vol) was warmedto approximately 40° C. before adding the solution of8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.The mixture was stirred for 1 hour. During this time a solidprecipitated. The solvent was exchanged to CPME (approximately 8volumes) via reduced pressure distillation and the resulting suspensionstirred at 15° C. for 2 hours. The resulting solid was collected anddried at 45° C. under vacuum to provide crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2. The crystalline form was confirmed by PXRD analysis using theInstrument Methods described in Example 77.

Example 72 Preparation of crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2

Methyl4-[(Z)-2-ethoxyethenyl]-2-[2-fluoro-4-(methylsulfanyl)anilino]-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate,prepared as described in Example 71, Step 1 (2.0 g, 5.0 mmol) wascombined with 2-Methyltetrahydrofuran (20 volumes) and heated toapproximately 75° C. O-(2-(tert-butoxy)ethyl)hydroxylamine hydrochloride(1.3 eq, 6.50 mmol) was added to the mixture and the resulting mixturecooled to 20° C. Borane-pyridine complex (2.0 eq, 10.0 mmol) was addedas well as HCl in cyclopentyl methyl ether (CPME) (3M, 1.3 eq, 6.5mmol). The resulting mixture was stirred at 60° C. After stirringovernight, the mixture was cooled to 20° C., quenched with HCl (1M, 12mL). During this time, some bubbling and exotherm was observed. Afterthe reaction appeared to be complete, (˜5 minutes), the organic layerwas separated, washed with water (8 vol), and concentrated toapproximately 6 volumes via reduced pressure distillation. At this time,phosphoric acid (14.6M, 15 eq, 75.0 mmol) was added and the mixturestirred at 65° C. After stirring overnight, the mixture was cooled to20° C. and diluted with 2-methyltetrahydrofuran (7 volumes). A solutionof potassium hydroxide (11.5M, 20 eq) was added slowly. After stirringfor approximately 10 minutes, the layers were allowed to separate. Theorganic phase was washed with water (10 mL) before concentrating toapproximately 5 vol with reduced pressure distillation. The mixture wasthen heated up to 50° C. with and cyclopentyl methyl ether (2 vol)added. Seeds of crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2 (prepared as described in Example 71) were added at this time andthe resulting suspension cooled to 30° C. Solvent was exchanged tocyclopentyl methyl ether (approximately 9 volumes) via reduced pressuredistillation and the resulting suspension stirred at 40° C. for 1 hour.The mixture was cooled to 20° C. and stirred overnight. The resultingsolid was collected and washed with cyclopentyl methyl ether (4 volumes)and dried at 45° C. under vacuum to provide crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2. FIG. 2 depicts a powder X-ray diffraction pattern of crystallineanhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2 prepared according to this method, wherein the PXRD analysis wasconducted using the Instrument Methods described in Example 77.

Example 73 Preparation of seed crystals of crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3

Into a reactor was charged 2-methyltetrahydrofuran (20 volumes) ofmethyl4-[(Z)-2-ethoxyethenyl]-2-[2-fluoro-4-(methylsulfanyl)anilino]-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate,prepared as described in Example 71, Step 1 (5.0 kg; limitingreagent—all subsequent chargers based on this quantity), andO-(2-(tert-butoxy)ethyl)hydroxylamine hydrochloride (2.89 kg). Thesubsequent mixture was heated to 70° C. After the reaction was complete,the mixture was cooled to 20° C. and borane-pyridine (8M, 3.3 L) and HClin CPME (3M, 5.95 L) were added. The mixture was stirred at 20° C. Afterreduction was complete, the mixture was heated to 65° C. untilcyclization was complete. The reaction was cooled to 25° C., quenchedwith 1 M HCl (6 volumes) and washed with water (8 volumes). The organiclayer was concentrated to approximately 7 volumes. Phosphoric acid (13.1L) was charged and the resulting mixture heated to 60° C. untildeprotection was complete. The reaction with quenched with aqueous 45wt. % potassium hydroxide (22.8 L) dissolved in water (7 volumes) andthen washed with water (10 volumes). The organic was then filteredthrough a spec free filter. At a reaction temperature of 45° C., theorganic layer was concentrated to approximately 5 volumes and the watercontent measured with KF and adjusted to 5% water. Approximately 5% ofthe reaction mixture was removed and cooled to 20° C. affording aslurry, which was charged back to the reactor to seed the bulk. Afterstirring at 45° C. for three hours, the solvent was exchanged to CPME (8volumes) with between 5%-10% residual 2-MeTHF and KF of at least 0.5%.The reaction was then cooled to 15° C. over 3 hours and granulated for 8hours. The solids were collected and the washed with CPME (4 volumes).The solids were dried under vacuum to remove residual solvent. Thedrying process dehydrated the solid form to crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2. The resulting crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2 was then rehydrated in an oven (without vacuum) at 25-40° C. withtrays of water. The final solids became rehydrated to afford crystallinemonohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3 based on KF and PXRD analysis, wherein the PXRD analysis wasconducted using the Instrument Methods described in Example 77.

Example 74 Preparation of crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3 by Seeded Crystallization

Into a reactor was charged 2-MeTHF (6 volumes, 19 L) and water (0.27volumes, 0.87 L). Amorphous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,prepared according to Example 6, Method A or B (3.22 kg) was added andthe mixture heated to 65° C. During this time the solid dissolved and asolution was obtained. The mixture was cooled to 45° C. and the seeds ofcrystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3 as prepared in Example 73 were added. The mixture was stirred forapproximately 2 hours before slowly charging heptane (4 volumes) over 3hours. The mixture was cooled to 15° C. over 4 hours and stirred for atleast 2 hours. The solids were then collected and washed with 50:50heptane/2-MeTHF (4 volumes, 13 L). The solids were dried under vacuum toremove residual solvent. The drying process dehydrated the solid form tocrystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2. The resulting crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2 was then rehydrated in an oven (without vacuum) at 25-40° C. withtrays of water. After sitting in the oven for 8-24 hours, the solid hadrehydrated afford crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3 based on KF and PXRD analysis.

Example 75 Alternative Preparation of Crystalline Monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3

Amorphous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,prepared according to Example 6, Method A or B (404.88 mg) was combinedwith 2-propanol (1.50 mL, 3.7 Volumes) with magnetic stirring. Themixture was heated to 60° C. During this time, the mixture becamehomogenous, dark red and all solids dissolved. Water (2.5 mL, 6.2volumes) was added and the mixture allowed to cool to room temperatureand stir for 2 hours. During this time, a solid precipitated, themixture became quite thick. A larger stir bar was added at this time. Analiquot was taken for analysis with PXRD and the mixture was allowed tostir at room temperature. After stirring overnight, the solid wascollected and washed with 9:1 2-propanol:water (2×1.00 mL). Dried inpreheated oven at 50° C. with pan of water for 3 hours followed bysitting overnight uncapped on the bench. 371 mg, 91% recovery. Theisolated material was determined to be crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3 by PXRD analysis. FIG. 3 depicts a powder X-ray diffractionpattern of crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3 prepared according to this method, wherein the PXRD analysis wasconducted using the Instrument Methods described in Example 77.

Example 76 Moisture Sorption Analysis of Crystalline Monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3

Water sorption and desorption studies were conducted on automated vaporsorption analyzer (TA instruments Q5000 SA). The microbalance wascalibrated using a 100 mg standard weight. The relative humidity sensorwas calibrated at 5.0, 11.3, 32.8, 52.8, 75.3, and 84.3% RH (25° C.)using saturated salt solutions. Approximately 10-20 mg of crystallinemonohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3 was placed in the quartz sample holder and dried at ≤3% relativehumidity (RH) at 25° C. The RH was then progressively increased from 0%to 40% RH in increments of 5% followed by a decrease to a final RH of 0%in 5% RH increments. A maximum equilibration time of 120 minutes wasused for all steps. The weight gain at each of the % RH steps is basedon the weight after the initial drying step at 0% RH. The weight changeof the sample was not used to assess equilibrium and instead all stepswere programed to be 120 minutes. After data collection was complete,data analysis was performed using commercially available TA universalanalysis software and Microsoft Excel.

FIG. 5 depicts the sorption isotherm of crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3. As shown in FIG. 5 , crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3 dehydrates to crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2 at 15% RH and 25° C. as confirmed by PXRD analysis. Crystallineanhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2 rehydrates to crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3 at 25% RH at 25° C. as confirmed by PXRD analysis. Based on thisanalysis, crystalline anhydrous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2 is stable below 10% RH at 25° C., and crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3 is stable above 30% RH at 25° C.

Example 77 General Procedures for Solid Form Analyses by PXRD InstrumentMethods

Powder X-ray diffraction analysis of anhydrous crystalline8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 1, anhydrous crystalline8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 2, and amorphous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dioneForm 4 was conducted using a Rigaku MiniFlex 6G Diffractometer equippedwith a Cu radiation source. The sample was prepared using a silicon lowbackground sample holder (2 mm×0.5 mm well). Diffracted radiation wasdetected by a D/teX Ultra2 detector. The X-ray tube voltage andamperages were set to 40 kV and 15 mA respectively. Data was collectedin the Miniflex goniometer at the Cu wavelength from 3.0 to 45.0°2-Theta using a step with of 0.02° and a step speed of 2.00°/minute. Theincident slit box was set to 1.25° and the length limiting slit was setat 10 mm. The sample was rotated at 10 RPM during collection. The datawas exported to *txt file using Rigaku software SmartLab Studio II andanalyzed with EVA diffract plus software. As shown by Example 76,anhydrous Form 2 rehydrates to monohydrate Form 3 at 25% RH at 25° C.Therefore, the analysis of anhydrous Form 2 by PXRD was conducted atapproximately 25° C. and at relative humidity below 10%.

Powder X-ray diffraction analysis of crystalline monohydrate8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione,Form 3 was conducted using a Bruker AXS D8 Endeavor diffractometerequipped with a Cu radiation source. The divergence slit was set at 10mm continuous illumination. Diffracted radiation was detected by aPSD-Lynx Eye detector, with the detector PSD opening set at 4.11degrees. The X-ray tube voltage and amperage were set to 40 kV and 40 mArespectively. Data was collected at the Cu wavelength (CuKā=1.5418 Δ) inthe Theta-Theta goniometer from 3.0 to 40.0 degrees 2-Theta. A step sizeof 0.02 degrees and a step time of 0.3 second was used. The antiscatterscreen was set to a fixed distance of 1.5 mm. Samples were rotatedduring data collection. Samples were prepared by placing them in asilicon low background sample holder and rotated during collection. Datawere collected using Bruker DIFFRAC Plus software and analysis wasperformed by EVA diffract plus software. The PXRD data file was notprocessed prior to peak searching. Using the peak search algorithm inthe EVA software, peaks selected with a threshold value of 1 were usedto make preliminary peak assignments. To ensure validity, adjustmentswere manually made; the output of automated assignments was visuallychecked, and peak positions were adjusted to the peak maximum. Peakswith relative intensity of ≥3% were generally chosen. Typically, thepeaks which were not resolved or were consistent with noise were notselected. A typical error associated with the peak position from PXRDstated in USP up to +/−0.2° 2-Theta (USP-941). As shown by Example 76,monohydrate Form 3 dehydrates to anhydrous Form 2 at 15% RH and 25° C.Accordingly, the PXRD analysis of monohydrate Form 3 with PXRD wasconducted at approximately 25° C. and at relative humidity above 30%.

PXRD Peak Picking Parameters

Using the peak search algorithm in the EVA software, peaks selected witha threshold value of 1 were used to make preliminary peak assignments.To ensure validity, adjustments were manually made; the output ofautomated assignments was visually checked, and peak positions wereadjusted to the peak maximum. Peaks with relative intensity of 3% weregenerally chosen. Typically, the peaks which were not resolved or wereconsistent with noise were not selected. A typical error associated withthe peak position from PXRD stated in USP up to +/−0.2° 2-Theta(USP-941).

It will be apparent to those skilled in the art that variousmodifications and variations may be made in the present inventionwithout departing from the scope or spirit of the invention. Otherembodiments of the invention will be apparent to those skilled in theart from consideration of the specification and practice of theinvention disclosed herein. It is intended that the specification andexamples be considered as exemplary only, with a true scope and spiritof the invention being indicated by the following claims.

All references cited herein, including patents, patent applications,papers, textbooks, and the like, and the references cited therein, tothe extent that they are not already, are hereby incorporated byreference in their entireties. In the event that one or more of theincorporated literature and similar materials differs from orcontradicts this application, including but not limited to definedterms, term usage, described techniques, or the like, this applicationcontrols.

Incorporated by reference herein in the entirety for all purposes is thecontent of U.S. Provisional Patent Application No. 63/168,456 filed Mar.31, 2021 and U.S. Provisional Patent Application No. 63/309,346 filedFeb. 11, 2022.

We claim:
 1. A method of treating a MEK-associated tumor, the methodcomprising: administering to a subject in need thereof a therapeuticallyeffective amount of a crystalline form of8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.2. The method of claim 1, where the crystalline form of8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dioneis a monohydrate or anhydrous.
 3. The method of claim 1, where thecrystalline form of8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dioneis anhydrous and has a PXRD pattern comprising peaks at 5.0, 8.7, 9.3,10.8, 14.5, 15.3, 18.8, and 20.5 degrees 2-theta (±0.2 degrees 2-theta).4. The method of claim 1, where the crystalline form of8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dioneis anhydrous and has a PXRD pattern comprising peaks at 7.1, 9.4, 12.4,12.8, 14.3, 15.6, 16.4, 17.4, 18.5, 18.9, 19.5, 19.9, 21.1, 21.4, 23.2,23.7, 24.8, 25.6, 27.6, 30.3, 33.2, 33.5, and 37.5 degrees 2-theta (±0.2degrees 2-theta).
 5. The method of claim 1, where the crystalline formof8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dioneis a monohydrate and has a PXRD pattern comprising peaks at 13.7, 18.0,and 18.3 degrees 2-theta (±0.2 degrees 2-theta).
 6. A method of treatinga MEK-associated tumor, the method comprising: administering to asubject in need thereof a therapeutically effective amount of amorphous8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.7. The method of claim 3, wherein the MEK-associated tumor has a BRAFV600 mutation selected from V600E, V600K, V600D, V600R and V600S.
 8. Themethod of claim 4, wherein the MEK-associated tumor has a BRAF V600mutation selected from V600E, V600K, V600D, V600R and V600S.
 9. Themethod of claim 5, wherein the MEK-associated tumor has a BRAF V600mutation selected from V600E, V600K, V600D, V600R and V600S.
 10. Themethod of claim 6, wherein the MEK-associated tumor has a BRAF V600mutation selected from V600E, V600K, V600D, V600R and V600S.
 11. Themethod of claim 3, wherein the MEK-associated tumor is a BRAF wild-typetumor.
 12. The method of claim 4, wherein the MEK-associated tumor is aBRAF wild-type tumor.
 13. The method of claim 5, wherein theMEK-associated tumor is a BRAF wild-type tumor.
 14. The method of claim6, wherein the MEK-associated tumor is a BRAF wild-type tumor.
 15. Themethod of claim 3, wherein the MEK-associated tumor is a CNS tumor. 16.The method of claim 4, wherein the MEK-associated tumor is a CNS tumor.17. The method of claim 5, wherein the MEK-associated tumor is a CNStumor.
 18. The method of claim 6, wherein the MEK-associated tumor is aCNS tumor.
 19. The method of claim 3, wherein the MEK-associated tumorhas a BRAF fusion.
 20. The method of claim 4, wherein the MEK-associatedtumor has a BRAF fusion.
 21. The method of claim 5, wherein theMEK-associated tumor has a BRAF fusion.
 22. The method of claim 6,wherein the MEK-associated tumor has a BRAF fusion.